CN104487440B

CN104487440B - The substituted Imidazopyridazine of amino

Info

Publication number: CN104487440B
Application number: CN201380017395.XA
Authority: CN
Inventors: K·艾斯; F·普勒; L·措恩; V·舒尔策; D·苏兹勒; P·利瑙; A·哈格巴尔特; K·彼得森; U·伯默
Original assignee: Bayer Pharma AG
Current assignee: Bayer Pharma AG
Priority date: 2012-03-29
Filing date: 2013-03-27
Publication date: 2016-11-23
Anticipated expiration: 2033-03-27
Also published as: US20150087631A1; ES2663609T3; JP2015511617A; HK1206009A1; CN104487440A; EP2858993B1; EP2858993A1; US9777004B2; CA2868673A1; WO2013144189A1; JP6173426B2

Description

The substituted Imidazopyridazine of amino

The present invention relates to the substituted Imidazopyridazine compounds of amino of as described herein and defined logical formula (I), system For the method for described compound, for preparing the midbody compound of described compound, comprising the drug regimen of described compound Thing and combination and described compound are used for preparing treatment or prevention disease as single medicament or with other active ingredient combinations The purposes of the pharmaceutical composition of (particularly hyperproliferative disorders and/or angiogenesis disorders).

Background of invention

The present invention relates to suppress MKNK1 kinases (also referred to map kinase interaction kinases, Mnk1) and MKNK2 kinases (also Be referred to as map kinase interaction kinases, Mnk2) compound.People MKNK includes one group by two kinds of gene (gene symbol: MKNK1 And MKNK2) four kinds of albumen being encoded by alternative splicing.B-type lacks the map kinase binding domain being positioned at C-end.MKNK1 Similar with the catalytic domain of MKNK2, and in subdomain VII, comprise exclusive DFD (Asp-Phe-Asp) motif, it is at it Its protein kinase is usually DFG (Asp-Phe-Gly) and is considered to change ATP combination [Jauch et al., Structure 13,1559-1568,2005 and Jauch et al., EMBO J25,4020-4032,2006].MKNK1a combines ERK and p38MAP Kinases and being activated by them, but is not activated by JNK1.MKNK2a combines ERK and is only activated by it.MKNK1b is at all conditions Under there is low activity, MKNK2b has Basal activity [Buxade M et al., the Frontiers unrelated with ERK or p38MAP kinases In Bioscience 5359-5374,2008 May 1].

It is proved MKNK phosphorylation eukaryotic initiation factor 4E (eIF4E), heterogeneous nRNA-associated proteins A1 (hnRNP A1), many pyrimidine track associated proteins dependency splicing factor (PSF), Cytoplasm phospholipase A2 (cPLA2) and Sprouty 2 (hSPRY2) [Buxade M et al., Frontiers in Bioscience 5359-5374,2008 May 1].

EIF4E is the oncogene being amplified in many cancers, and as shown in KO-mice study [Konicek et al., Cell Cycle 7:16,2466-2471,2008；Ueda et al., Mol Cell Biol 24,6539-6549,2004], only by MKNK protein phosphorylation.EIF4E plays pivotal role in terms of making cellular mRNA translation.EIF4E combines 5 ' ends of cell mRNA The 7-methylguanosine cap at place, and they are delivered to a ribosome part as eIF4F complex, this complex also includes EIF4G and eIF4A.Although all of mRNA of capping needs eIF4E to translate, but mRNA pond depends on rising singularly EIF4E activity is to translate.These so-called " weak mRNA " are typically due to their length and 5 ' UTR region of complexity and by more Less efficiently translate, and they encode the albumen all played a significant role at all aspects of malignant tumor, including VEGF, FGF- 2, c-Myc, cyclin D1, survivin, BCL-2, MCL-1, MMP-9, heparitinase etc..The expression of eIF4E and merit Can raise in multiple human cancer, and the most relevant to progression of disease [Konicek et al., Cell Cycle 7:16, 2466-2471,2008]。

MKNK1 and MKNK2 is only known kinases of phosphorylation eIF4E at Ser209.Overall translation rate is not subject to The impact of eIF4E phosphorylation, it has been shown that eIF4E phosphorylation promotes finally to make " weak mRNA " more effectively to be translated Polysome forms (i.e. multiple ribosome on single mRNA) [Buxade M et al., Frontiers in Bioscience 5359-5374,2008 May 1].Or, MKNK protein phosphorylation eIF4E can promote that eIF4E discharges from 5 ' caps so that 48S complex can be mobile along " weak mRNA ", to position start codon [Blagden SP and Willis AE, Nat Rev Clin Oncol.8(5):280-91,2011].Therefore, poor pre-predictive of Patients with Non-small-cell Lung of the eIF4E phosphorylation of increase Afterwards [Yoshizawa et al., Clin Cancer Res.16 (1): 240-8,2010].Other data show, MKNK1 is at carcinogenic work Function affect in, because the constitutively activate MKNK1 in mouse embryo fibroblasts (rather than kinase dead MKNK1) process LAN promote tumor formed [Chrestensen C.A. et al., Genes Cells 12,1133-1140, 2007].Additionally, the MKNK protein phosphorylation increased is relevant with active process LAN in breast carcinoma to HER2 [Chrestensen, C.A. et al., J.Biol.Chem.282,4243-4252,2007].By E μ-Myc transgenic Hematopoietic Stem Cell is for producing in blastomogenic model in mice, and the MKNK1 of composition activation (rather than kinase dead) also promotes tumor Growth.When analyzing the eIF4E with S209D sudden change, obtain similar result.S209D sudden change simulates at MKNK1 phosphoric acid Change the phosphorylation of site.On the contrary, eIF4E can not reduce tumor growth [Wendel HG et al., Genes by phosphorylation form Dev.21(24):3232-7,2007].Block the selectivity MKNK inhibitor of eIF4E phosphorylation in vitro induction of apoptosis And inhibit propagation and the soft agar growth of cancerous cell.This inhibitor further suppress experimental B16 melanoma Lung metastases Outgrowth and the growth of subcutaneous HCT116 colon cancer xenograft tumor, and do not affect body weight [Konicek et al., Cancer Res.71(5):1849-57,2011].In a word, cell proliferation can be promoted via the eIF4E phosphorylation of MKNK protein active and deposit Live, and most important for vicious transformation.The suppression of MKNK activity can provide the cancer treatment method being prone to control.

WO 2007/025540 A2 (Bayer Schering Pharma AG) relates to substituted imidazo [1,2-b] and rattles away Piperazine, it is as inhibitors of kinases, particularly PKC (Protein kinase C) inhibitor, particularly PKC theta inhibitors.

WO 2007/025090 A2 (Kalypsis, Inc.) relates to heterocyclic compound, and it can be used as mitogen and activates The inhibitor of protein kinase (MAPK)/extracellular signal-regulated kinase (Erk) kinases (being abbreviated as " MEK ").Especially, WO 2007/025090 A2 particularly relates to imidazo [1,2-b] pyridazine.

WO 2007/013673 A1 (Astellas Pharma Inc.) relates to annelated heterocycles, and it is as lymphocyte egg The inhibitor of white tyrosine kinase (being abbreviated as " LCK ").Especially, WO 2007/013673 A1 particularly relates to imidazo [1,2- B] pyridazine.

WO 2007/147646 A1 (Bayer Schering Pharma AG) relates to oxo substituted imidazo [1,2- B] pyridazine, it is as inhibitors of kinases, particularly PKC (Protein kinase C) inhibitor, particularly PKC theta inhibitors.

WO 2008/025822 A1 (Cellzome (UK) Ltd.) relates to deriving as diazole the diazine of inhibitors of kinases Thing.Especially, WO 2008/025822 A1 particularly relates to imidazo [1,2-b] pyridazine, and it is used as inhibitors of kinases, particularly Can inducing T cell kinases (being abbreviated as " Itk ") inhibitor.

It is receptor associated sharp that WO 2008/030579 A2 (Biogen Idec MA Inc.) relates to il-1 (IL-1) The regulator of enzyme (being abbreviated as " IRAK ").Especially, WO 2008/030579 A2 particularly relates to imidazo [1,2-b] pyridazine.

WO 2008/058126 A2 (Supergen, Inc.) particularly relates to imidazo [1,2-b] pyridyl derivatives, and it is made For kinases inhibitor, particularly PIM inhibitors of kinases.

WO 2009/060197 A1(Centro Nacional de Investigaciones Oncologicas (CNIO)) relating to Imidazopyridazine, it is used as protein kinase (such as PIM family kinase) inhibitor.

US 4,408,047 (Merck&Co., Inc.) particularly relates to the miaow with 3-amino-2-OR-propoxyl group substituent group Azoles pyridazine, it has beta-adrenergic blockade activity.

WO 03/018020 A1 (Takeda Chemical Industries, Ltd.) relates to swashing for c-Jun N-end The inhibitor of enzyme, it comprises especially for the compound of imidazo [1,2-b] pyridazine.

WO 2008/052734 A1 (Novartis AG) relates to the heterocyclic compound as antiinflammatory.Especially, described Compound is especially imidazo [1,2-b] pyridazine.It is receptor-mediated by ALK-5 and/or ALK-4 that described compound can be used for treatment Disease, and also can be used for treatment by PI3K receptor, JAK-2 receptor and the receptor-mediated disease of TRK.

WO 2008/072682 A1 (Daiichi Sankyo Company, Limited) relates to imidazo [1,2-b] and rattles away Oxazine derivatives, it has the effect that suppression TNF-α produces, in the pathological model of inflammatory diseases and/or autoimmune disease Play a role.

WO 2008/079880 A1 (Alcon Research, Ltd.) relates to 6-aminooimidazole also [1,2-b] pyridazine and is similar to Thing, it is with acting on treatment glaucoma and the Rho-inhibitors of kinases of ocular hypertension.

WO 2009/091374 A2 (Amgen Inc.) relates to condensed heterocyclic derivates.Selected compound is the most pre- Prevent and treatment disease, such as hepatocyte growth factor (" HGF ") disease.

At J.Med.Chem., 2005,48, 7604-7614 is entitled " Structural Basis of Inhibitor Specificity of the Protooncogene Proviral Insertion Site in Moloney Murine Leukemia Virus (PIM-1) Kinase " article, and in particular disclose and grind for wherein said as inhibitor structure Imidazo [1,2-b] pyridazine studied carefully.

At J.Med.Chem., 2010,53, 6618-6628 is entitled " Discovery of Mitogen- Activated Protein Kinase-Interacting Kinase 1Inhibitors by a Comprehensive Fragment-Oriented Virtual Screening Approach " article, and disclose the most in Table 1 some make For being confirmed to be concrete imidazo [1,2-b] pyridazine of the compound of MKNK-1 inhibitor.

At Cancer Res, on March 1st, 2011,71, 1849-1857 is entitled " Therapeutic inhibition of MAP kinase interacting kinase blocks eukaryotic initiation factor 4E Phosphorylation and suppresses outgrowth of experimental lung mestastases " Article, and in particular disclose known antifungal cercosporamide (Cercosporamide) be MKNK1 inhibitor.

But, above-mentioned prior art is silent on such as described herein and definition and the hereinafter referred to as " change of the present invention Compound " the present invention as herein defined lead to specific substituted Imidazopyridazine compounds or its stereoisomerism of formula (I) Body, tautomer, N-oxide, hydrate, solvate or salt, or their mixture, or its pharmacological activity, institute State specific substituted Imidazopyridazine compounds the most such imidazo [1,2-b] pyridazinyl moieties, its:

-on its 3 it is:

Group；

-there is on its 6 the group of following structure:

Wherein:

-* represents the junction point of described group and molecule remainder,

-R1 represents the linear C being optionally substituted as defined herein₁-C₆-alkyl-, branched C₃-C₆-alkyl-or C₃-C₆-cycloalkyl-, and

-R5 represents:

Or:

Zero substituent group as herein defined；

Or:

The carbon atom of zero nitrogen-atoms being connected with it and R1 is collectively forming 3 yuan as herein defined and swells to 7 rings Amido；

Have now found that the compound of the present invention has astonishing and favourable character, and which constitute the base of the present invention Plinth.

Specifically, in fact it has surprisingly been found that the compound of the described present invention effectively suppresses MKNK-1 kinases and therefore can be used for Treatment or prevention by uncontrolled cell growth, breed and/or survive, unsuitable cellullar immunologic response or unsuitable carefully The disease that born of the same parents' inflammatory response causes, or be attended by uncontrolled cell growth, breed and/or survive, unsuitable cell is exempted from Epidemic disease response or the disease of unsuitable cellular inflammation response, especially, wherein said uncontrolled cell growth, propagation and/ Or survival, unsuitable cellullar immunologic response or unsuitable cellular inflammation response are kinase mediated by MKNK-1, such as blood Liquid tumor, solid tumor and/or their transfer, such as leukemia and myelodysplastic syndrome, malignant lymphoma, include brain Tumor and brain metastes swell at interior head and tumor colli, chest including non-fire power and small cell lung tumor Tumor, gastroenteric tumor, endocrine tumors, breast tumor and other gynecological tumor, include tumor of kidney, tumor of bladder and prostatitis adenoncus Tumor is in interior urologic neoplasms, cutaneous tumor and sarcoma, and/or their transfer.

Summary of the invention

According to first aspect, the present invention contains compound or its stereoisomer, tautomer, the N-oxygen of logical formula (I) Compound, hydrate, solvate or salt, or their mixture:

Wherein:

Represent:

Group；

Wherein * represents the junction point of described group and molecule remainder；And

R1 represents linear C₁-C₆-alkyl-, branched C₃-C₆-alkyl-or C₃-C₆-cycloalkyl, it is optionally selected from Following substituent group replacement once or independently of each other replaces repeatedly:

Halogen atom ,-CN, C₁-C₆-alkyl-, C₁-C₆-haloalkyl-, C₂-C₆-thiazolinyl-, C₂-C₆-alkynyl-, C₃-C₁₀- Cycloalkyl-, aryl-,-C (=O) NH₂,-C (=O) N (H) R ' ,-C (=O) N (R ') R " ,-C (=O) OH ,-C (=O) OR ' ,- NH₂,-NHR ' ,-N (R ') R " ,-N (H) C (=O) R ' ,-N (R ') C (=O) R ' ,-N (H) S (=O) R ' ,-N (R ') S (=O) R ' ,-N (H) S (=O)₂R ' ,-N (R ') S (=O)₂R ' ,-N=S (=O) (R ') R " ,-OH, C₁-C₆-alkoxyl-, C₁-C₆-halo Alkoxyl-,-OC (=O) R ' ,-OC (=O) NH₂,-OC (=O) NHR ' ,-OC (=O) N (R ') R " ,-SH, C₁-C₆-alkyl- S-,-S (=O) R ' ,-S (=O)₂R ' ,-S (=O)₂NH₂,-S (=O)₂NHR ' ,-S (=O)₂N(R’)R”；

R2 represents hydrogen atom；

R3 represents selected from following substituent group:

Halogen atom ,-CN, C₁-C₆-alkyl-, C₁-C₆-haloalkyl-, C₂-C₆-thiazolinyl-, C₂-C₆-alkynyl-,-C (=O) R ' ,-C (=O) NH₂,-C (=O) N (H) R ' ,-C (=O) N (R ') R " ,-NH₂,-NHR ' ,-N (R ') R " ,-N (H) C (=O) R ' ,- N (R ') C (=O) R ' ,-N (H) C (=O) NH₂,-N (H) C (=O) NHR ' ,-N (H) C (=O) N (R ') R " ,-N (R ') C (=O) NH₂,-N (R ') C (=O) NHR ' ,-N (R ') C (=O) N (R ') R " ,-N (H) C (=O) OR ' ,-N (R ') C (=O) OR ' ,-NO₂、- N (H) S (=O) R ' ,-N (R ') S (=O) R ' ,-N (H) S (=O)₂R ' ,-N (R ') S (=O)₂R ' ,-N=S (=O) (R ') R " ,- OH、C₁-C₆-alkoxyl-, C₁-C₆-halogenated alkoxy-,-OC (=O) R ' ,-SH, C₁-C₆-alkyl-S-,-S (=O) R ' ,-S (= O)₂R ' ,-S (=O)₂NH₂,-S (=O)₂NHR ' ,-S (=O)₂N (R ') R " ,-S (=O) (=NR ') R "；

R4 represents selected from following substituent group:

Hydrogen atom, halogen atom ,-CN, C₁-C₆-alkyl-, C₁-C₆-haloalkyl-, C₂-C₆-thiazolinyl-, C₂-C₆-alkynyl-, C₃-C₁₀-cycloalkyl-, 3 yuan to 10 yuan Heterocyclylalkyls-, optionally replaced once by R substituent or replace independently of each other repeatedly Aryl-；Optionally replaced once by R substituent or replace independently of each other heteroaryl repeatedly-；-C (=O) NH₂,-C (= O) N (H) R ' ,-C (=O) N (R ') R " ,-C (=O) OR ' ,-NH₂,-NHR ' ,-N (R ') R " ,-N (H) C (=O) R ' ,-N (R ') C (=O) R ' ,-N (H) C (=O) NH₂,-N (H) C (=O) NHR ' ,-N (H) C (=O) N (R ') R " ,-N (R ') C (=O) NH₂、-N (R ') C (=O) NHR ' ,-N (R ') C (=O) N (R ') R " ,-N (H) C (=O) OR ' ,-N (R ') C (=O) OR ' ,-NO₂、-N(H)S (=O) R ' ,-N (R ') S (=O) R ' ,-N (H) S (=O)₂R ' ,-N (R ') S (=O)₂R ' ,-N=S (=O) (R ') R " ,-OH, C₁- C₆-alkoxyl-, C₁-C₆-halogenated alkoxy-,-OC (=O) R ' ,-OC (=O) NH₂,-OC (=O) NHR ' ,-OC (=O) N (R ') R”、-SH、C₁-C₆-alkyl-S-,-S (=O) R ' ,-S (=O)₂R ' ,-S (=O)₂NH₂,-S (=O)₂NHR ' ,-S (=O)₂N (R ') R " ,-S (=O) (=NR ') R "；

R represents selected from following substituent group:

Halogen atom ,-CN, C₁-C₆-alkyl-, C₁-C₆-haloalkyl-, C₂-C₆-thiazolinyl-, C₂-C₆-alkynyl-, C₃-C₁₀- Cycloalkyl-, 3 yuan to 10 yuan Heterocyclylalkyls-, aryl-, heteroaryl-,-C (=O) R ' ,-C (=O) NH₂,-C (=O) N (H) R ' ,- C (=O) N (R ') R " ,-C (=O) OR ' ,-NH₂,-NHR ' ,-N (R ') R " ,-N (H) C (=O) R ' ,-N (R ') C (=O) R ' ,-N (H) C (=O) NH₂,-N (H) C (=O) NHR ' ,-N (H) C (=O) N (R ') R " ,-N (R ') C (=O) NH₂,-N (R ') C (=O) NHR ' ,-N (R ') C (=O) N (R ') R " ,-N (H) C (=O) OR ' ,-N (R ') C (=O) OR ' ,-NO₂,-N (H) S (=O) R ' ,-N (R ') S (=O) R ' ,-N (H) S (=O)₂R ' ,-N (R ') S (=O)₂R ' ,-N=S (=O) (R ') R " ,-OH, C₁-C₆-alcoxyl Base-, C₁-C₆-halogenated alkoxy-,-OC (=O) R ' ,-OC (=O) NH₂,-OC (=O) NHR ' ,-OC (=O) N (R ') R " ,-SH, C₁-C₆-alkyl-S-,-S (=O) R ' ,-S (=O)₂R ' ,-S (=O)₂NH₂,-S (=O)₂NHR ' ,-S (=O)₂N(R’)R”、-S (=O) (=NR ') R "；

R ' and R " represent independently of each other selected from following substituent group:

C₁-C₆-alkyl-, C₃-C₁₀-cycloalkyl-, C₁-C₆-haloalkyl；

R5 represents:

Or:

-selected from following substituent group: C₁-C₆-alkyl-, C₁-C₆-haloalkyl-, C₂-C₆-thiazolinyl-, C₂-C₆-alkynyl-, C₃-C₁₀-cycloalkyl-, C₃-C₁₀-cycloalkyl-C₁-C₆-alkyl-, aryl-,-C (=O) NH₂,-C (=O) N (H) R ' ,-C (=O) N (R ') R " ,-S (=O) R ' ,-S (=O)₂R’；

Or:

The carbon atom of-the nitrogen-atoms that is connected with it and R1 is collectively forming 3 yuan and swells amido to 7 rings, its optionally by Replace selected from following substituent group:

N represents the integer of 0,1,2,3,4 or 5.

Term mentioned in this article preferably has a following implication:

Term " halogen atom ", " halo-" or " halogen-" is understood to mean that fluorine atom, chlorine atom, bromine atoms or iodine are former Son, preferably fluorine atom, chlorine atom, bromine atoms or atomic iodine.

Term " C₁-C₆-alkyl " it is interpreted as preferably representing that there is the linear or branched of 1,2,3,4,5 or 6 carbon atoms Saturated monovalent hydrocarbon, such as methyl, ethyl, propyl group, butyl, amyl group, hexyl, isopropyl, isobutyl group, sec-butyl, the tert-butyl group, Isopentyl, 2-methyl butyl, 1-methyl butyl, 1-ethyl propyl, 1,2-dimethyl propyl, neopentyl, 1,1-dimethyl propyl, 4-methyl amyl, 3-methyl amyl, 2-methyl amyl, 1-methyl amyl, 2-ethyl-butyl, 1-ethyl-butyl, 3,3-dimethyl Butyl, 2,2-dimethylbutyl, 1,1-dimethylbutyl, 2,3-dimethylbutyl, 1,3-dimethylbutyl or 1,2-dimethyl Butyl or their isomer.Especially, described group has 1,2,3 or 4 carbon atom (" C₁-C₄-alkyl "), such as methyl, Ethyl, propyl group, butyl, isopropyl, isobutyl group, sec-butyl, the tert-butyl group, more particularly, it is former that described group has 1,2 or 3 carbon Son (" C₁-C₃-alkyl "), such as methyl, ethyl, n-pro-pyl or isopropyl.

Term " halo-C₁-C₆-alkyl " it is interpreted as preferably representing that wherein one or more hydrogen atoms are with identical or different Mode replaced the linear or branched saturated monovalent hydrocarbon of (i.e. between halogen atom separate) by halogen atom, wherein Term " C₁-C₆-alkyl " as defined above.Especially, described halogen atom is F.Described halo-C₁-C₆-alkyl be such as- CF₃、-CHF₂、-CH₂F、-CF₂CF₃Or-CH₂CF₃。

Term " C₁-C₆-alkoxyl " it is interpreted as the linear or branched saturated monovalent hydrocarbon of preferred expression-O-alkyl Base, wherein term " alkyl " is as defined above, such as methoxyl group, ethyoxyl, positive propoxy, isopropoxy, n-butoxy, isobutyl Epoxide, tert-butoxy, sec-butoxy, amoxy, isoamoxy or positive hexyloxy, or their isomer.

Term " halo-C₁-C₆-alkoxyl " it is interpreted as preferably representing that wherein one or more hydrogen atoms are with identical or not The most linear or branched saturated monovalence C that same mode is replaced by halogen atom₁-C₆-alkoxyl.Especially, Described halogen atom is F.Described halo-C₁-C₆-alkoxyl is such as-OCF₃、-OCHF₂、-OCH₂F、-OCF₂CF₃Or- OCH₂CF₃。

Term " C₁-C₆-alkoxy-C₁-C₆-alkyl " it is interpreted as preferably representing that wherein one or more hydrogen atoms are with phase Same or different mode is by C as defined above₁-C₆The most linear or branched saturated one that-alkoxyl replaces Valency alkyl or their isomer, wherein term " C₁-C₆-alkyl " as defined above, such as methoxyalkyl, ethyoxyl alkyl, Allyloxyalkyl, isopropoxy alkyl, butoxy alkyl, isobutoxy alkyl, tert-butoxy alkyls, sec-butoxy alkyl, penta Epoxide alkyl, isoamoxy alkyl, hexyloxy alkyl.

Term " halo-C₁-C₆-alkoxy-C₁-C₆-alkyl " it is interpreted as preferably representing wherein one or more hydrogen atoms The most linear or branched saturated monovalence C replaced by halogen atom in the way of identical or different₁-C₆-alcoxyl Base-C₁-C₆-alkyl.Especially, described halogen atom is F.Described halo-C₁-C₆-alkoxy-C₁-C₆-alkyl be such as- CH₂CH₂OCF₃、-CH₂CH₂OCHF₂、-CH₂CH₂OCH₂F、-CH₂CH₂OCF₂CF₃Or-CH₂CH₂OCH₂CF₃。

Term " C₂-C₆-thiazolinyl " it is interpreted as preferably representing linear or branched monovalent hydrocarbon, it comprises one or many Individual double bond and there are 2,3,4,5 or 6 carbon atoms, particularly 2 or 3 carbon atom (" C₂-C₃-thiazolinyl "), it should be appreciated that in institute Stating in the case of thiazolinyl comprises more than one double bond, described double bond can be separated from each other or be conjugated.Described thiazolinyl is such as ethylene Base, pi-allyl, (E)-2-methyl ethylene, (Z)-2-methyl ethylene, high allyl, (E)-but-2-ene base, (Z)-butyl-2- Thiazolinyl, (E)-but-1-ene base, (Z)-but-1-ene base, amyl-4-thiazolinyl, (E)-amyl-3-thiazolinyl, (Z)-amyl-3-thiazolinyl, (E)- Amyl-2-thiazolinyl, (Z)-amyl-2-thiazolinyl, (E)-amyl-1-thiazolinyl, (Z)-amyl-1-thiazolinyl, hex-5-thiazolinyl, (E)-hex-4-thiazolinyl, (Z)-hex-4-thiazolinyl, (E)-hex-3-thiazolinyl, (Z)-hex-3-thiazolinyl, (E)-hex-2-thiazolinyl, (Z)-hex-2-thiazolinyl, (E)-hex- 1-thiazolinyl, (Z)-hex-1-thiazolinyl, isopropenyl, 2-methyl-prop-2-thiazolinyl, 1-methyl-prop-2-thiazolinyl, 2-methyl-prop-1-alkene Base, (E)-1-methyl-prop-1-thiazolinyl, (Z)-1-methyl-prop-1-thiazolinyl, 3-methyl butyl-3-thiazolinyl, 2-methyl butyl-3-thiazolinyl, 1- Methyl butyl-3-thiazolinyl, 3-methyl but-2-ene base, (E)-2-methyl but-2-ene base, (Z)-2-methyl but-2-ene base, (E)-1- Methyl but-2-ene base, (Z)-1-methyl but-2-ene base, (E)-3-methyl but-1-ene base, (Z)-3-methyl but-1-ene base, (E)-2-methyl but-1-ene base, (Z)-2-methyl but-1-ene base, (E)-1-methyl but-1-ene base, (Z)-1-methyl but-1-ene Base, 1,1-dimethyl propylene-2-thiazolinyl, 1-ethyl acrylate-1-thiazolinyl, 1-propyl ethylene base, 1-isopropyl-ethylene base, 4-methylpent- 4-thiazolinyl, 3-methylpent-4-thiazolinyl, 2-methylpent-4-thiazolinyl, 1-methylpent-4-thiazolinyl, 4-methylpent-3-thiazolinyl, (E)-3- Methylpent-3-thiazolinyl, (Z)-3-methylpent-3-thiazolinyl, (E)-2-methylpent-3-thiazolinyl, (Z)-2-methylpent-3-thiazolinyl, (E)-1-methylpent-3-thiazolinyl, (Z)-1-methylpent-3-thiazolinyl, (E)-4-methylpent-2-thiazolinyl, (Z)-4-methylpent-2-alkene Base, (E)-3-methylpent-2-thiazolinyl, (Z)-3-methylpent-2-thiazolinyl, (E)-2-methylpent-2-thiazolinyl, (Z)-2-methylpent- 2-thiazolinyl, (E)-1-methylpent-2-thiazolinyl, (Z)-1-methylpent-2-thiazolinyl, (E)-4-methylpent-1-thiazolinyl, (Z)-4-methyl Amyl-1-thiazolinyl, (E)-3-methylpent-1-thiazolinyl, (Z)-3-methylpent-1-thiazolinyl, (E)-2-methylpent-1-thiazolinyl, (Z)-2- Methylpent-1-thiazolinyl, (E)-1-methylpent-1-thiazolinyl, (Z)-1-methylpent-1-thiazolinyl, 3-ethyl butyl-3-thiazolinyl, 2-ethyl Butyl-3-thiazolinyl, 1-ethyl butyl-3-thiazolinyl, (E)-3-ethyl but-2-ene base, (Z)-3-ethyl but-2-ene base, (E)-2-ethyl But-2-ene base, (Z)-2-ethyl but-2-ene base, (E)-1-ethyl but-2-ene base, (Z)-1-ethyl but-2-ene base, (E)-3- Ethyl but-1-ene base, (Z)-3-ethyl but-1-ene base, 2-ethyl but-1-ene base, (E)-1-ethyl but-1-ene base, (Z)-1- Ethyl but-1-ene base, 2-propyl group acrylate-2-thiazolinyl, 1-propyl group acrylate-2-thiazolinyl, 2-isopropyl acrylate-2-thiazolinyl, 1-isopropyl acrylate-2- Thiazolinyl, (E)-2-propyl group acrylate-1-thiazolinyl, (Z)-2-propyl group acrylate-1-thiazolinyl, (E)-1-propyl group acrylate-1-thiazolinyl, (Z)-1-propyl group Acrylate-1-thiazolinyl, (E)-2-isopropyl acrylate-1-thiazolinyl, (Z)-2-isopropyl acrylate-1-thiazolinyl, (E)-1-isopropyl acrylate-1-thiazolinyl, (Z)-1-isopropyl acrylate-1-thiazolinyl, (E)-3,3-dimethyl propylene-1-thiazolinyl, (Z)-3,3-dimethyl propylene-1-thiazolinyl, 1-(1,1- Dimethyl ethyl) vinyl, butyl-1,3-dialkylene, amyl-1,4-dialkylene, hex-1,5-dialkylene or methyl hexadienyl.Special Not, described group is vinyl or pi-allyl.

Term " C₂-C₆-alkynyl " it is interpreted as preferably representing linear or branched monovalent hydrocarbon, it comprises one or many Individual three keys and there are 2,3,4,5 or 6 carbon atoms, particularly 2 or 3 carbon atom (" C₂-C₃-alkynyl ").Described C₂-C₆-alkynes Base be such as acetenyl, acrylate-1-alkynyl, Propargyl, butyl-1-alkynyl, butyl-2-alkynyl, butyl-3-alkynyl, amyl-1-alkynyl, Amyl-2-alkynyl, amyl-3-alkynyl, amyl-4-alkynyl, hex-1-alkynyl, hex-2-alkynyl, hex-3-alkynyl, hex-4-alkynyl, hex-5-alkynes Base, 1-methyl Propargyl, 2-methyl butyl-3-alkynyl, 1-methyl butyl-3-alkynyl, 1-methyl butyl-2-alkynyl, 3-methyl butyl- 1-alkynyl, 1-ethyl Propargyl, 3-methylpent-4-alkynyl, 2-methylpent-4-alkynyl, 1-methylpent-4-alkynyl, 2-methyl Amyl-3-alkynyl, 1-methylpent-3-alkynyl, 4-methylpent-2-alkynyl, 1-methylpent-2-alkynyl, 4-methylpent-1-alkynyl, 3- Methylpent-1-alkynyl, 2-ethyl butyl-3-alkynyl, 1-ethyl butyl-3-alkynyl, 1-ethyl butyl-2-alkynyl, 1-propyl group acrylate-2-alkynes Base, 1-isopropyl Propargyl, 2,2-dimethyl butyrate-3-alkynyl, 1,1-dimethyl butyrate-3-alkynyl, 1,1-dimethyl butyrate-2- Alkynyl or 3,3-dimethyl butyrate-1-alkynyl.Especially, described alkynyl is acetenyl, acrylate-1-alkynyl or Propargyl.

Term " C₃-C₁₀-cycloalkyl " be understood to mean that saturated monovalent monocyclic or dicyclo hydrocarbon ring, it has 3,4,5,6, 7,8,9 or 10 carbon atom (" C₃-C₁₀-cycloalkyl ").Described C₃-C₁₀-cycloalkyl is such as monocycle hydrocarbon ring such as cyclopropyl, ring fourth Base, cyclopenta, cyclohexyl, suberyl, ring octyl group, ring nonyl or ring decyl, or dicyclo hydrocarbon ring such as perhydro pentalene Or decahydronaphthalene naphthalene nucleus (perhydropentalenylene).The most described ring comprises 3,4,5 or 6 carbon atom (" C₃-C₆-ring Alkyl ").

Term " C₄-C₁₀-cycloalkenyl group " be interpreted as preferably representing monovalent monocyclic or dicyclo hydrocarbon ring, it comprises 4,5,6,7,8, 9 or 10 carbon atoms and 1,2,3 or 4 double bonds being conjugated or be not conjugated, as long as the size of described cyclenes basic ring allows.Such as, Described C₄-C₁₀-cycloalkenyl group is the monocycle hydrocarbon ring of such as cyclobutane base, cyclopentenyl or cyclohexenyl group, or dicyclo hydrocarbon ring, example As:

Term " 3 yuan to 10 yuan Heterocyclylalkyls " is understood to mean that saturated monovalent monocyclic or dicyclo hydrocarbon ring, it has 2, 3,4,5,6,7,8 or 9 carbon atoms and one or more selected from C (=O), O, S, S (=O), S (=O)₂、NR^aContaining hetero atom Group, wherein R^aRepresent hydrogen atom or C₁-C₆-alkyl-or halo-C₁-C₆-alkyl-；Described Heterocyclylalkyl can be by described Any one or nitrogen-atoms (if present) in carbon atom are connected with the remainder of molecule.

Especially, described 3 yuan to 10 yuan Heterocyclylalkyls can have 2,3,4 or 5 carbon atoms and one or more on Stating containing heteroatomic group (" 3 yuan to 6 yuan Heterocyclylalkyls "), more particularly, it is former that described Heterocyclylalkyl can have 4 or 5 carbon Sub and one or more above-mentioned containing heteroatomic group (" 5 yuan to 6 yuan Heterocyclylalkyls ").

Especially, described Heterocyclylalkyl can be such as but not limited to: 4 rings, such as azetidinyl (azetidinyl), oxetanyl (oxetanyl)；5 rings, such as tetrahydrofuran base, dioxa cyclopentenyl (dioxolinyl), pyrrolidinyl, imidazolidinyl, pyrazolidinyl, pyrrolinyl；Or 6 rings, as THP trtrahydropyranyl, piperidyl, Morpholinyl, dithiane base (dithianyl), tetrahydro-1,4-thiazine base, piperazinyl or trithiane base (trithianyl)；Or 7 rings, such as two Azepan basic ring.Optionally, described Heterocyclylalkyl can be benzo-fused.

Described heterocyclic radical can be dicyclo, such as but not limited to 5,5 rings, as hexahydro cyclopentano [c] pyrroles-2 (1H)- Basic ring, or 5,6 yuan of dicyclos, such as hexahydropyrrolo also [1,2-a] pyrazine-2 (1H)-basic ring.

Being as previously mentioned, described nitrogen atom ring can be that part is undersaturated, and i.e. it can comprise one or more double Key, such as but not limited to 2,5-dihydro-1H-pyrrole radicals, 4H-[1,3,4] thiadiazine base, 4,5-dihydro oxazolyl or 4H-[Isosorbide-5-Nitrae] Thiazine basic ring, or, it can be benzo-fused, such as but not limited to dihydro-isoquinoline basic ring.

Term " 4 yuan to 10 yuan heterocycloalkenyl " is understood to mean that undersaturated monovalent monocyclic or dicyclo hydrocarbon ring, and it has 3,4,5,6,7,8 or 9 carbon atoms and one or more selected from C (=O), O, S, S (=O), S (=O)₂、NR^aContaining hetero atom Group, wherein R^aRepresent hydrogen atom or C₁-C₆-alkyl or halo-C₁-C₆-alkyl；Described heterocycloalkenyl can pass through described carbon Any one or nitrogen-atoms (if present) in atom are connected with the remainder of molecule.The example of described heterocycloalkenyl can To comprise one or more double bond, the double aziridinyl (3H-diazirinyl) of such as 4H-pyranose, 2H-pyranose, 3H-, 2, 5-dihydro-1H-pyrrole radicals, [1,3]-dioxa cyclopentenyl ([1,3] dioxolyl), 4H-[1,3,4] thiadiazine base, 2,5- Dihydrofuran base, 2,3 dihydro furan base, 2,5-dihydro-thiophene base (dihydrothiophenyl), 2,3-dihydro-thiophene base, 4, 5-dihydro oxazolyl or 4H-[Isosorbide-5-Nitrae] thiazinyl, or it can be benzo-fused.

Term " 3 yuan to the 7 rings amidos that swell " is understood to mean that selected from following group:

Wherein:

Rx represents hydrogen atom, C₁-C₆-alkyl-or halo-C₁-C₆-alkyl-；And

* the junction point of described group and molecule remainder is represented.

Term " aryl " is interpreted as preferably representing the monovalence virtue with 6,7,8,9,10,11,12,13 or 14 carbon atoms Fragrance or the monocycle of partial aromatic, dicyclo or tricyctic hydrocarbon ring (" C₆-C₁₄-aryl "), particularly there is the ring of 6 carbon atoms (“C₆-aryl "), such as phenyl；Or xenyl, or there is the ring (" C of 9 carbon atoms₉-aryl "), such as indanyl or Indenyl, or there is the ring (" C of 10 carbon atoms₁₀-aryl "), such as tetrahydro naphthyl, dihydro naphthyl or naphthyl, or There is the ring (" C of 13 carbon atoms₁₃-aryl "), such as fluorenyl, or there is the ring (" C of 14 carbon atoms₁₄-aryl "), Such as anthryl.

Term " heteroaryl " is interpreted as preferably representing such monovalent monocyclic, dicyclo or tricyclic aromatic ring system, and it has 5,6,7,8,9,10,11,12,13 or 14 annular atomses (" 5 yuan to 14 yuan heteroaryls "), particularly 5 or 6 or 9 or 10 carbon are former Son, and it comprises at least one hetero atom (described hetero atom is such as oxygen, nitrogen or sulfur) that can be identical or different, and, separately Outer can be benzo-fused at each occurrence.Especially, heteroaryl selected from thienyl, furyl, pyrrole radicals, oxazolyl, Thiazolyl, imidazole radicals, pyrazolyl, isoxazolyl, isothiazolyl, di azoly, triazolyl, thiadiazolyl group, thiophene-4H-pyrazolyl Etc. and their benzo derivative, such as benzofuranyl, benzothienyl, benzothiazole (thia-4H-pyrazolyl) Base, benzisoxa oxazolyl, benzimidazolyl, benzotriazole base, indazolyl, indyl, isoindolyl etc.；Or pyridine radicals, pyridazine Base, pyrimidine radicals, pyrazinyl, triazine radical etc., and their benzo derivative, such as quinolyl, quinazolyl, isoquinolyl Deng；Or azocine base (azocinyl), indolizine base, purine radicals etc. and their benzo derivative；Or cinnolines base, phthalazinyl, Quinazolyl, quinoxalinyl, naphthyridinyl (naphthpyridinyl), pteridyl, carbazyl, acridinyl, phenazinyl, phenothiazine Base, phenazinyl, ton base or oxepin base (oxepinyl) etc..

And except as otherwise noted, described heteroaryl or inferior heteroaryl include its all possible isomery shape generally speaking Formula, such as its position isomer.Therefore, for some illustrative limiting examples, term pyridine radicals or pyridylidene bag Include pyridine-2-base, sub-pyridine-2-base, pyridin-3-yl, sub-pyridin-3-yl, pyridin-4-yl and sub-pyridin-4-yl；Or, art Language thienyl or sub-thienyl include thiophene-2-base, sub-thiophene-2-base, thiene-3-yl and sub-thiene-3-yl.

As used the most in the whole text, such as at " C₁-C₆-alkyl ", " C₁-C₆-haloalkyl ", " C₁-C₆-alkoxyl " or " C₁- C₆-halogenated alkoxy " definition linguistic context in use term " C₁-C₆" it is understood to mean that the carbon with 1-6 limited quantity Atom, the alkyl of i.e. 1,2,3,4,5 or 6 carbon atoms.Should also be understood that described term " C₁-C₆" be to be understood as included in therein Anyon scope, such as C₁-C₆、C₂-C₅、C₃-C₄、C₁-C₂、C₁-C₃、C₁-C₄、C₁-C₅；Particularly C₁-C₂、C₁-C₃、C₁-C₄、C₁- C₅、C₁-C₆；More particularly C₁-C₄；At " C₁-C₆-haloalkyl " or " C₁-C₆-halogenated alkoxy " in the case of, more particularly It is C₁-C₂。

Similarly, as used herein, use the most in the whole text, such as at " C₂-C₆-thiazolinyl " and " C₂-C₆-alkynyl " Definition linguistic context in use term " C₂-C₆" it is understood to mean that the carbon atom with 2-6 limited quantity, i.e. 2,3,4,5 Or the alkenyl or alkynyl of 6 carbon atoms.Should also be understood that described term " C₂-C₆" should be interpreted that the anyon model being contained therein Enclose, such as C₂-C₆、C₃-C₅、C₃-C₄、C₂-C₃、C₂-C₄、C₂-C₅；Particularly C₂-C₃。

It addition, as used herein, use the most in the whole text, such as at " C₃-C₆-cycloalkyl " definition linguistic context in Term " the C used₃-C₆" it is understood to mean that the carbon atom with 3-6 limited quantity, the ring of i.e. 3,4,5 or 6 carbon atoms Alkyl.Should also be understood that described term " C₃-C₆" should be interpreted that the anyon scope being contained therein, such as C₃-C₆、C₄-C₅、C₃- C₅、C₃-C₄、C₄-C₆、C₅-C₆；Particularly C₃-C₆。

Term " substituted " refers to that the one or more hydrogen on specified atom are replaced by the selection from pointed group, Condition is to form stable compound not less than specified atom normal atom valency in the current situation and described replacement. The combination of substituent group and/or variable is only only permission when this combination forms stable compound.

Term " optionally replacement " refers to optionally be replaced by specific group, atomic group or part.

The substituent group of ring system refers to the substituent group being connected with aromatics or non-aromatic ring system, and the most described substituent group replaces described ring Fasten available hydrogen.

The term that uses (such as in the definition of the substituent group of the general formula compound of the present invention) herein is " once or many Secondary " be understood to mean that once, twice, three times, four times or five times, the most once, twice, three times or four times, more particularly Once, twice or thrice, the most once or twice.

Present invention additionally comprises all applicable isotopic variations of the compound of the present invention.The coordination of the compound of the present invention Element variant is defined as such compound, and at least one of which atom is had same atoms ordinal number but atomic mass is different from certainly So in boundary, the atom of atomic mass that is common or that mainly find replaces.The isotope in the compound of the present invention can be incorporated into Example include the isotope of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, chlorine, bromine and iodine, the most such as²H (deuterium),³H (tritium),¹³C、¹⁴C 、¹⁵N、¹⁷O、¹⁸O、³²P、³³P、³³S、³⁴S、³⁵S、³⁶S、¹⁸F、³⁶Cl、⁸²Br、¹²³I、¹²⁴I、¹²⁹I and¹³¹I.The compound of the present invention Some isotopic variations, wherein introduce one or more such as³H or¹⁴C radioisotopic those, can be used for medicine Thing and/or substrate tissue distribution research.Due to easily prepared and detectability, the most tritiated isotope and carbon-14 (i.e.¹⁴C) isotope.Some can be provided by controlling that higher metabolic stability causes additionally, replace with isotope (such as deuterium) Treat benefit, such as Half-life in vivo to increase or dose requirements reduction, and be preferred the most in some cases.The change of the present invention The isotopic variations of compound generally can be prepared by conventional method well known by persons skilled in the art, such as by shown method Or use the suitable isotopic variations of the reagent being suitable for prepare by the preparation method described in below example.

When word used herein: during the plural form of compound, salt, polymorph, hydrate, solvate etc., Also single compound, salt, polymorph, isomer, hydrate, solvate etc. are meant.

" stable compound " or " stable structure " refers to the most powerful, it is possible to stand to be separated to from reactant mixture Purity and be configured to the compound of effective therapeutic agent.

The compound of the present invention can comprise one or more asymmetric center, depends on the position of desired different substituents Put and character.Asymmetric carbon atom can exist by (R) or (S) configuration, only during an asymmetric center, produces raceme mixing Thing, during containing multiple asymmetric center, obtains non-enantiomer mixture.In some cases, due to around particular key Blocked rotation there is likely to be unsymmetry, and such as this center key connects two aromatic rings being replaced of specific compound.

The compound of the present invention can comprise sulphur atom, and it can be asymmetric, the asymmetric Asia of following structure Sulfone or sulphoxide imine (sulphoximine):

Such as

Wherein * represents the atom can being combined with molecule remainder.

Substituent group on ring can exist with cis or trans form.Expect that this type of configuration all of (includes enantiomerism Body and diastereomer) all it is included in the scope of the present invention.

Preferably compound be produce more desirable bioactive those.The separation of the compound of the present invention, pure Or partially purified isomer and stereoisomer or racemic mixture or non-enantiomer mixture are the most all included in In the scope of the invention.The purification of this type of material and separation can be realized by standard technique known in the art.

Optical isomer can be obtained according to conventional methods, such as by using the acid of optically-active by resolving racemic mixtures Or alkali forms diastereomeric salt, or by forming covalent diastereomeric.The example of suitable acid be tartaric acid, two Acetyl group tartaric acid, ditoluoyltartaric and camphorsulfonic acid.The mixture of diastereomer can physics based on them And/or chemical differences, it is separated into their list by methods known in the art (such as by chromatography or fractional crystallization) The diastereomer of one.Then, from the diastereomeric salt separated, discharge alkali or the acid of optically-active.Another kind of different dividing Method from optical isomer relates to using chiral chromatography (such as chirality under conditions of carrying out or do not carry out conventional derivation HPLC column), it can be through optimum selection to separate enantiomer substantially.The chirality HPLC column being suitable for is to be given birth to by Daicel Produce, such as Chiracel OD and Chiracel OJ etc., all of all can select routinely.Also carrying out or can not derive Enzyme process is used to separate under conditions of change.Similarly, the rotation of the present invention can be obtained by using the chirality synthesis of the raw material of optically-active Optical compounds.

In order to be defined between different types of isomer, with reference to IUPAC Rules Section E (Pure Appl Chem 45,11-30,1976)。

The present invention includes all possible stereoisomer of the compound of the present invention, and it is single stereoisomers or institute State arbitrary proportion the most mixed of stereoisomer (such as R-isomer or S-isomer, or E-isomer or Z-isomer) The form of compound.Can be realized by the art methods (such as chromatography, the most such as chiral chromatography) being arbitrarily suitable for The separation of the single stereoisomers (the most single enantiomer or single diastereomer) of the compound of the present invention.

It addition, the compound of the present invention can be presented in tautomer.Such as, pyrazolyl is arbitrarily comprised as miscellaneous The compound of the present invention of aryl such as can be presented in 1H tautomer or 2H tautomer, or even with arbitrarily Presented in the mixture of the two tautomer of amount, or triazolyl such as can be mutual with 1H tautomer, 2H Presented in tautomeric or 4H tautomer, or even mixed with described 1H, 2H and 4H tautomer of any amount Presented in compound, it may be assumed that

The present invention includes all possible tautomer of the compound of the present invention, and it is single tautomer or institute State the form of any mixture of the arbitrary proportion of tautomer.

It addition, the compound of the present invention can be presented in N-oxide, it is defined as in the compound of the present invention At least one nitrogen is oxidized.The present invention includes this type of possible N-oxides all.

The invention still further relates to the useful form of compound as disclosed herein, such as metabolite, hydrate, solvate, Prodrug, salt, particularly pharmaceutically acceptable salt and co-precipitation thing.

The compound of the present invention can be presented in hydrate or solvate, and wherein the compound of the present invention comprises work For the polar solvent of the structural element of described compound lattice, the most such as water, methanol or ethanol.Polar solvent particularly water Amount can exist with stoichiometric proportion or non-stoichiometric.In the case of stoichiometric solvates (such as hydrate), May be respectively half (hemi-) solvate or hydrate, (half (semi-)) solvate or hydrate, a solvate or Hydrate, sesquialter solvate or hydrate, two solvates or hydrate, three solvates or hydrate, four solvates Or hydrate, five solvates or hydrate etc..The present invention includes this type of hydrates all or solvate.

It addition, the compound of the present invention can exist in a free form, such as with free alkali, free acid or zwitterionic shape Formula, or exist in a salt form.Described salt can be any salt, and it can be organic addition salts or inorganic addition salts, particularly medicine Any pharmaceutically acceptable organic addition salts conventional in or inorganic addition salts.

Term " pharmaceutically acceptable salt " refers to the relative nontoxic of the compound of the present invention, mineral acid or organic acid addition Salt.For example, with reference to S.M.Berge et al., " Pharmaceutical Salts, " J.Pharm.Sci.1977,66,1-19.

The pharmaceutically acceptable salt being suitable for of the compound of the present invention can be such as to have nitrogen-atoms in chain or ring The acid-addition salts of compound with enough present invention of alkalescence, the acid-addition salts such as formed with following mineral acid: such as Hydrochloric acid, hydrobromic acid, hydroiodic acid, sulphuric acid, pyrosulfuric acid (bisulfuric acid), phosphoric acid or nitric acid, or with following organic acid shape Become acid-addition salts: such as formic acid, acetic acid, acetoacetic acid, acetone acid, trifluoroacetic acid, propanoic acid, butanoic acid, caproic acid, enanthic acid, 11 Alkanoic acid, lauric acid, benzoic acid, salicylic acid, 2-(4-hydroxy benzoyl) benzoic acid, dextrocamphoric acid., cinnamic acid, Pentamethylene. propanoic acid, Didextrose acid (digluconic acid), 3-hydroxy-2-naphthoic acid, nicotinic acid, flutter acid, pectinic acid, persulfuric acid, 3-phenyl third Acid, picric acid, pivalic acid, 2-ethylenehydrinsulfonic acid, itaconic acid, sulfamic acid, trifluoromethanesulfonic acid, lauryl sulphate acid, ethyl sulfonic acid, Benzenesulfonic acid, p-methyl benzenesulfonic acid, methanesulfonic acid, 2-LOMAR PWA EINECS 246-676-2, naphthalenedisulfonic acid, camphorsulfonic acid, citric acid, tartaric acid, stearic acid, breast Acid, oxalic acid, malonic acid, succinic acid, malic acid, adipic acid, alginic acid, maleic acid, fumaric acid, D-gluconic acid, mandelic acid, Vitamin C Acid, glucoheptose, phosphoglycerol, aspartic acid, sulfosalicylic acid, hemisulfic acid (hemisulfuric acid) or Hydrogen thiocyanate.

It addition, the pharmaceutically acceptable salt that the another kind with the compound of the enough acid present invention is suitable for is alkali gold Belong to salt (such as sodium salt or potassium salt), alkali salt (such as calcium salt or magnesium salt), ammonium salt, or with provide the acceptable sun of physiology The salt that the organic base of ion is formed, the salt such as formed with following material: N-METHYL-ALPHA-L-GLUCOSAMINE, dimethyl glycosamine, ethyl Portugal Osamine, lysine, dicyclohexylamine, 1,6-hexamethylene diamine, ethanolamine, glycosamine, sarcosine, serinol, trihydroxy methyl amino Methane, amino-propanediol, sovak-alkali, 1-amino-2,3,4-butantriol.It addition, Basic nitrogen-containing groups can be with following reagent season Ammonium: elementary alkyl halide, such as methyl, ethyl, propyl group and butyl chloride compound, bromide and iodide；Sulphuric acid dialkyl group Ester, such as dimethyl sulfate, dithyl sulfate, dibutyl sulfate and diamyl sulfates；Long chain halide, such as decyl, Laurel Base, myristyl and stearyl chlorides, bromide and iodide；Aralkyl halide such as benzyl and phenylethyl bromide etc..

Skilled persons will also appreciate that, the acid-addition salts of compound required for protection can pass through multiple known formula Any one in method makes described compound prepare with suitable mineral acid or organic acid reaction.Or, the acidity of the present invention Alkali metal salt and the alkali salt of compound make the compound of the present invention and suitable alkali reaction by various known methods Prepare.

The present invention includes all possible salt of the compound of the present invention, and it is any of single salt form or described salt The form of any mixture of ratio.

Terms used herein " internal hydrolyzable ester " is understood to mean that the change of the present invention comprising carboxyl or hydroxyl The internal hydrolyzable ester of compound, such as, can be hydrolyzed in human body or animal body thus produce pharmaceutically may be used of parent acid or alcohol The ester accepted.The pharmaceutically acceptable ester being suitable for for carboxyl includes such as Arrcostab, cycloalkyl ester and is optionally substituted Phenylalkyl ester, particularly benzyl ester, C₁-C₆Alkoxy methyl ester (such as methoxymethyl ester), C₁-C₆Alkanoyloxy first Base ester (such as pivaloyloxymethyl ester, phthalidyl ester), C₃-C₈Cycloalkyloxy carbonyloxy group-C₁-C₆Arrcostab (such as 1-cyclohexyl Carbonyloxy group ethyl ester), 1,3-dioxole-2-carbonvlmethyl ester (1,3-dioxolen-2-onylmethylester) (such as 5-methyl isophthalic acid, 3-dioxole-2-carbonvlmethyl ester), and C₁-C₆-Cialkoxycarbonyloxyethyl esters (such as 1- Methoxycarbonyloxyethyl ester), and described ester can be formed on any carboxyl of the compound of the present invention.

The internal hydrolyzable ester of the compound of the present invention comprising hydroxyl includes inorganic acid ester (such as phosphate ester), [α] Acyloxyalkyi ethers and related compound, described related compound breaks to form parent hydroxyl due to the internal hydrolysis of described ester Base.The example of [α] acyloxyalkyi ethers includes acetoxymethoxy (acetoxymethoxy) and 2,2-dimethyl propionyl Oxymethoxy ether (2,2-dimethylpropionyloxymethoxy).With the group that hydroxyl forms internal hydrolyzable ester Selection include alkanoyl, benzoyl, phenyl acetyl and substituted benzoyl and phenyl acetyl, alkoxy carbonyl group (with Form alkyl carbonate), dialkyl carbamoyl and N-(di-alkyaminoethyl group)-N-alkyl-carbamoyl be (to be formed Carbamate), dialkylaminoacetyl and carboxyacetyl.The present invention includes this type of esters all.

It addition, the present invention includes all possible crystal form or the polymorph of the compound of the present invention, it can be single The mixture of the arbitrary proportion of one polymorph or more than one polymorph.

The second embodiment according to described first aspect, the present invention contains compound or its solid of above-mentioned logical formula (I) Isomer, tautomer, N-oxide, hydrate, solvate or salt, or their mixture, wherein:

Represent:

Group；

R2 represents hydrogen atom；

R3 represents selected from following substituent group:

Halogen atom ,-CN, C₁-C₆-alkyl-, C₁-C₆-haloalkyl-,-OH, C₁-C₆-alkoxyl-, C₁-C₆-alkyl halide Epoxide-；

R4 represents selected from following substituent group:

R represents selected from following substituent group:

C₁-C₆-alkyl-, C₃-C₁₀-cycloalkyl-, C₁-C₆-haloalkyl；

R5 represents:

Or:

N represents the integer of 0,1,2,3,4 or 5.

The 3rd embodiment according to described first aspect, the present invention contains compound or its solid of above-mentioned logical formula (I) Isomer, tautomer, N-oxide, hydrate, solvate or salt, or their mixture, wherein:

Represent:

Group；

R2 represents hydrogen atom；

R3 represents selected from following substituent group:

R4 represents selected from following substituent group:

Hydrogen atom, halogen atom ,-CN, C₁-C₆-alkyl-, C₁-C₆-haloalkyl, C₃-C₁₀-cycloalkyl-, aryl-, miscellaneous Aryl-；

C₁-C₆-alkyl-, C₃-C₁₀-cycloalkyl-, C₁-C₆-haloalkyl；

R5 represents:

Or:

The carbon atom of-the nitrogen-atoms that is connected with it and R1 is collectively forming 3 yuan and swells amido to 7 rings；

N represents the integer of 0 or 1.

The 4th embodiment according to described first aspect, the present invention contains compound or its solid of above-mentioned logical formula (I) Isomer, tautomer, N-oxide, hydrate, solvate or salt, or their mixture, wherein:

Represent:

Group；

R1 represents linear C₁-C₅-alkyl-, branched C₃-C₅-alkyl-or C₄-C₆-cycloalkyl, it is optionally selected from Following substituent group replacement once or independently of each other replaces repeatedly:

C₁-C₆-alkyl-or aryl-；

R2 represents hydrogen atom；

R3 represents selected from following substituent group:

R4 represents selected from following substituent group:

C₁-C₆-alkyl-, C₃-C₁₀-cycloalkyl-, C₁-C₆-haloalkyl；

R5 represents:

Or:

N represents the integer of 0 or 1.

The 5th embodiment according to described first aspect, the present invention contains compound or its solid of above-mentioned logical formula (I) Isomer, tautomer, N-oxide, hydrate, solvate or salt, or their mixture, wherein:

Represent:

Group；

R1 represents linear C₁-C₅-alkyl-, it is optionally substituted base and replaces once, and described substituent group is:

Aryl-；

R2 represents hydrogen atom；

R3 represents selected from following substituent group:

Halogen atom, C₁-C₆-alkoxyl-；

R4 represents hydrogen atom；

R5 represents:

Or:

-selected from following substituent group: C₁-C₆-alkyl-, C₃-C₁₀-cycloalkyl-, C₃-C₁₀-cycloalkyl-C₁-C₆-alkyl-；

Or:

N represents the integer of 0 or 1.

According to the another embodiment of above-mentioned aspect, the present invention relates to the compound of formula (I), wherein:

Represent:

Group；

Wherein * represents the junction point of described group and molecule remainder.

Halogen atom ,-CN, C₁-C₆-alkyl-, C₁-C₆-haloalkyl-, C₂-C₆-thiazolinyl-, C₂-C₆-alkynyl-, C₃-C₁₀- Cycloalkyl-, aryl-,-C (=O) NH₂,-C (=O) N (H) R ' ,-C (=O) N (R ') R " ,-C (=O) OH ,-C (=O) OR ' ,- NH₂,-NHR ' ,-N (R ') R " ,-N (H) C (=O) R ' ,-N (R ') C (=O) R ' ,-N (H) S (=O) R ' ,-N (R ') S (=O) R ' ,-N (H) S (=O)₂R ' ,-N (R ') S (=O)₂R ' ,-N=S (=O) (R ') R " ,-OH, C₁-C₆-alkoxyl-, C₁-C₆-halo Alkoxyl-,-OC (=O) R ' ,-OC (=O) NH₂,-OC (=O) NHR ' ,-OC (=O) N (R ') R " ,-SH, C₁-C₆-alkyl- S-,-S (=O) R ' ,-S (=O)₂R ' ,-S (=O)₂NH₂,-S (=O)₂NHR ' ,-S (=O)₂N(R’)R”。

R2 represents hydrogen atom.

R3 represents selected from following substituent group:

Halogen atom ,-CN, C₁-C₆-alkyl-, C₁-C₆-haloalkyl-, C₂-C₆-thiazolinyl-, C₂-C₆-alkynyl-,-C (=O) R ' ,-C (=O) NH₂,-C (=O) N (H) R ' ,-C (=O) N (R ') R " ,-NH₂,-NHR ' ,-N (R ') R " ,-N (H) C (=O) R ' ,- N (R ') C (=O) R ' ,-N (H) C (=O) NH₂,-N (H) C (=O) NHR ' ,-N (H) C (=O) N (R ') R " ,-N (R ') C (=O) NH₂,-N (R ') C (=O) NHR ' ,-N (R ') C (=O) N (R ') R " ,-N (H) C (=O) OR ' ,-N (R ') C (=O) OR ' ,-NO₂、- N (H) S (=O) R ' ,-N (R ') S (=O) R ' ,-N (H) S (=O)₂R ' ,-N (R ') S (=O)₂R ' ,-N=S (=O) (R ') R " ,- OH、C₁-C₆-alkoxyl-, C₁-C₆-halogenated alkoxy-,-OC (=O) R ' ,-SH, C₁-C₆-alkyl-S-,-S (=O) R ' ,-S (= O)₂R ' ,-S (=O)₂NH₂,-S (=O)₂NHR ' ,-S (=O)₂N (R ') R " ,-S (=O) (=NR ') R ".

R4 represents selected from following substituent group:

Hydrogen atom, halogen atom ,-CN, C₁-C₆-alkyl-, C₁-C₆-haloalkyl-, C₂-C₆-thiazolinyl-, C₂-C₆-alkynyl-, C₃-C₁₀-cycloalkyl-, 3 yuan to 10 yuan Heterocyclylalkyls-, optionally replaced once by R substituent or replace independently of each other repeatedly Aryl-；Optionally replaced once by R substituent or replace independently of each other heteroaryl repeatedly-；-C (=O) NH₂,-C (= O) N (H) R ' ,-C (=O) N (R ') R " ,-C (=O) OR ' ,-NH₂,-NHR ' ,-N (R ') R " ,-N (H) C (=O) R ' ,-N (R ') C (=O) R ' ,-N (H) C (=O) NH₂,-N (H) C (=O) NHR ' ,-N (H) C (=O) N (R ') R " ,-N (R ') C (=O) NH₂、-N (R ') C (=O) NHR ' ,-N (R ') C (=O) N (R ') R " ,-N (H) C (=O) OR ' ,-N (R ') C (=O) OR ' ,-NO₂、-N(H)S (=O) R ' ,-N (R ') S (=O) R ' ,-N (H) S (=O)₂R ' ,-N (R ') S (=O)₂R ' ,-N=S (=O) (R ') R " ,-OH, C₁- C₆-alkoxyl-, C₁-C₆-halogenated alkoxy-,-OC (=O) R ' ,-OC (=O) NH₂,-OC (=O) NHR ' ,-OC (=O) N (R ') R”、-SH、C₁-C₆-alkyl-S-,-S (=O) R ' ,-S (=O)₂R ' ,-S (=O)₂NH₂,-S (=O)₂NHR ' ,-S (=O)₂N (R ') R " ,-S (=O) (=NR ') R ".

R represents selected from following substituent group:

Halogen atom ,-CN, C₁-C₆-alkyl-, C₁-C₆-haloalkyl-, C₂-C₆-thiazolinyl-, C₂-C₆-alkynyl-, C₃-C₁₀- Cycloalkyl-, 3 yuan to 10 yuan Heterocyclylalkyls-, aryl-, heteroaryl-,-C (=O) R ' ,-C (=O) NH₂,-C (=O) N (H) R ' ,- C (=O) N (R ') R " ,-C (=O) OR ' ,-NH₂,-NHR ' ,-N (R ') R " ,-N (H) C (=O) R ' ,-N (R ') C (=O) R ' ,-N (H) C (=O) NH₂,-N (H) C (=O) NHR ' ,-N (H) C (=O) N (R ') R " ,-N (R ') C (=O) NH₂,-N (R ') C (=O) NHR ' ,-N (R ') C (=O) N (R ') R " ,-N (H) C (=O) OR ' ,-N (R ') C (=O) OR ' ,-NO₂,-N (H) S (=O) R ' ,-N (R ') S (=O) R ' ,-N (H) S (=O)₂R ' ,-N (R ') S (=O)₂R ' ,-N=S (=O) (R ') R " ,-OH, C₁-C₆-alcoxyl Base-, C₁-C₆-halogenated alkoxy-,-OC (=O) R ' ,-OC (=O) NH₂,-OC (=O) NHR ' ,-OC (=O) N (R ') R " ,-SH, C₁-C₆-alkyl-S-,-S (=O) R ' ,-S (=O)₂R ' ,-S (=O)₂NH₂,-S (=O)₂NHR ' ,-S (=O)₂N(R’)R”、-S (=O) (=NR ') R ".

C₁-C₆-alkyl-, C₃-C₁₀-cycloalkyl-, C₁-C₆-haloalkyl.

R5 represents:

-selected from following substituent group: C₁-C₆-alkyl-, C₁-C₆-haloalkyl-, C₂-C₆-thiazolinyl-, C₂-C₆-alkynyl-, C₃-C₁₀-cycloalkyl-, C₃-C₁₀-cycloalkyl-C₁-C₆-alkyl-, aryl-,-C (=O) NH₂,-C (=O) N (H) R ' ,-C (=O) N (R ') R " ,-S (=O) R ' ,-S (=O)₂R’。

R5 represents:

N represents the integer of 0,1,2,3,4 or 5.

R3 represents selected from following substituent group:

R4 represents selected from following substituent group:

Hydrogen atom, halogen atom ,-CN, C₁-C₆-alkyl-, C₁-C₆-haloalkyl, C₃-C₁₀-cycloalkyl-, aryl-, miscellaneous Aryl-.

R5 represents:

The carbon atom of-the nitrogen-atoms that is connected with it and R1 is collectively forming 3 yuan and swells amido to 7 rings.

N represents the integer of 0 or 1.

C₁-C₆-alkyl-or aryl-.

Aryl-.

R3 represents selected from following substituent group:

Halogen atom, C₁-C₆-alkoxyl-.

R4 represents hydrogen atom.

C₁-C₆-alkyl-, C₃-C₁₀-cycloalkyl-.

R5 represents:

-selected from following substituent group: C₁-C₆-alkyl-, C₃-C₁₀-cycloalkyl-, C₃-C₁₀-cycloalkyl-C₁-C₆-alkyl-.

N represents the integer of 0.

N represents the integer of 1.

According to the another embodiment of above-mentioned aspect, the present invention relates to the compound of the formula (I) of any of the above-described embodiment Stereoisomer, tautomer, N-oxide, hydrate, solvate or salt, or the form of their mixture.

In should be appreciated that any embodiment or the aspect of the compound of the logical formula (I) that the present invention relates to the invention described above Any sub-combination.

More particularly, the present invention contains the compound of the logical formula (I) disclosed in Examples below part.

According on the other hand, the present invention contains the method for the compound of the preparation present invention, and described method includes real herein Test the step described in part.

On the other hand according to, the present invention contains leads to the compound of formula (I) and (retouches the most in this article for preparing the present invention In the method stated) midbody compound.Especially, the present invention contains the compound of logical formula V:

Wherein the compound of A, R2, R3, R4 and n as above mutual-through type (I) is defined, and X represents leaving group, such as halogen Element atom (such as chlorine atom, bromine atoms or atomic iodine), or perfluoroalkyl sulfonate ester base is (such as trifluoromethane sulfonic acid ester group or nine fluorine Butyl sulfonic acid ester group).

According on the other hand, the present invention contains the midbody compound of logical formula V for preparing formula as defined above (I) purposes of compound:

Wherein the compound of A, R2, R3, R4 and n as above mutual-through type (I) is defined, and X represents leaving group, such as halogen Element atom (such as chlorine atom, bromine atoms or atomic iodine), or perfluoroalkyl sulfonate ester base (such as trifluoromethane sulfonic acid ester group).

Experimental section

Following table lists the abbreviation used in this joint and embodiment part.

Abbreviation	Implication
		DMF	Dimethylformamide
DMSO	Dimethyl sulfoxide

THF	Oxolane
		NMR	Nuclear magnetic resonance, NMR
MS	Mass spectrum
		R_t	Retention time
HPLC,LC	High performance liquid chromatography
		H	Hour
min	Minute

The synthesis (general introduction) of compound

Can be as with the compound describing the preparation present invention in lower part.Route 1 described below and operation illustrate The general synthetic routes of the compound of the logical formula (I) of the present invention and the most restrictive.Those skilled in the art are aobvious and easy See the order that can change the conversion illustrated in route 1 in many ways.Therefore, the conversion illustrated in route 1 Order is the most restrictive.It addition, substituent R 1, R2, R3, R4, R5 can be realized before or after the conversion illustrated With the change of any one in A.These are modified can be such as to introduce protection group, crack protection group, exchange, reduce or oxidation official's energy Group, halogenation, metallize, replace or well known by persons skilled in the art other reaction.These conversions include that introducing makes substituent group enter Those of the degree of functionality of one step change convert.Suitable protection group and their introducing and to be cracked into those skilled in the art public Know (see, e.g. T.W.Greene and P.G.M.Wuts in Protective Groups in Organic Synthesis, 3rd edition, Wiley 1999).Subsequent paragraph describes concrete example.Further it is possible that can carry out two or more Individual continuous print step and between described step, do not carry out post processing, such as " one kettle way " reaction, this is those skilled in the art Known.

Route 1:

Wherein A, R1, R2, R3, R4, R5 and n are as defined above, and X and Y represents leaving group, such as halogen atom (such as chlorine atom, bromine atoms or atomic iodine), or perfluoroalkyl sulfonate ester base is (such as trifluoromethane sulfonic acid ester group or nine fluorine butyl sulphurs Perester radical).

In the first step, the compound (i.e. with the dichloro-pyridazine of applicable X substituent group) of formula A and ammonia can be raised Under temperature and pressure react, with obtain Formula B compound [similar with WO200733080, by its with its entirety quote addition Literary composition is as reference].

In second step, by the compound of Formula B and such as 2-Chloro-1-ethanal two acetal or bromoacetaldehyde two aldolisation, to obtain Bicyclic ring system C [similar with DE102006029447, it to be quoted with its entirety and is added herein by reference].

Can be such as by using the bromo-butanimide of N-or the iodo-butanimide of N-by the compound bromination of formula C respectively Or iodate, it is achieved 3 activation of bicyclic system, to obtain the compound of general formula D.

In the 4th step, the available catalyzed coupling reaction being suitable for, use such as boric acid or stannane (stannane), it is achieved Group A-[R3]_nIntroducing, this generate general formula E compound.

The compound of general formula E is used as to introduce the center intermediate of the multiple side chain comprising alcohol functional group, and this generates formula (I) Imidazopyridazine-ether.Can be by such as using alkali (such as sodium hydride) to realize the introducing of side chain.According to the character of side chain, can Must can carry out these reactions at elevated temperatures.Have in the functional group being likely to must be introduced into may interfere with desired reaction There is the side chain of applicable protection group.

As illustrated by route 2, the 4th step of described step and the 5th step are the most interchangeable.

Route 2:

According to an embodiment, the present invention also relates to the method that the compound of formula (I) is led in preparation as defined above, described Method includes making the midbody compound of logical formula V:

Wherein the compound of A, R2, R3, R4 and n as above mutual-through type (I) is defined, and X represents leaving group, such as halogen Element atom (such as chlorine atom, bromine atoms or atomic iodine), or perfluoroalkyl sulfonate ester base is (such as trifluoromethane sulfonic acid ester group or nine fluorine Butyl sulfonic acid ester group),

The step reacted with the compound of logical formula (III):

Wherein the compound of R1 and R5 as above mutual-through type (I) is defined,

Thus obtain the compound of logical formula (I):

Wherein A, R1, R2, R3, R4, R5 and n are as defined above.

Common segment

ACD/Name Batch Version 12.01 is used to generate chemical name.

HPLC method:

Method 1:

Instrument: Waters Acquity UPLCMS ZQ4000；Post: Acquity UPLC BEH C181.7 μm, 50x2.1mm；Eluent A: water+0.05 volume % formic acid, eluent B: acetonitrile+0.05 volume % formic acid；Gradient: 0-1.6min 1-99%B, 1.6-2.0min 99%B；Flow velocity: 0.8mL/min；Temperature: 60 DEG C；Sample size: 2 μ L；DAD scans: 210- 400nm；ELSD

Method 2:

Instrument: Waters Acquity UPLCMS SQD3001；Post: Acquity UPLC BEH C181.7 μm, 50x2.1mm；Eluent A: water+0.1 volume % formic acid (95%), eluent B: acetonitrile；Gradient: 0-1.6min 1-99%B, 1.6-2.0min 99%B；Flow velocity: 0.8mL/min；Temperature: 60 DEG C；Sample size: 2 μ L；DAD scans: 210-400nm；ELSD

Method 3:

Instrument: Waters Acquity UPLCMS SQD；Post: Acquity UPLC BEH C181.7 μm, 50x2.1mm； Eluent A: water+0.05 volume % formic acid (95%), eluent B: acetonitrile+0.05 volume % formic acid (95%)；Gradient: 0- 1.6min 1-99%B, 1.6-2.0min 99%B；Flow velocity: 0.8mL/min；Temperature: 60 DEG C；Sample size: 2 μ L；DAD scans: 210-400nm；ELSD

Method 4 instrument: Waters Acquity UPLCMS SQD；Post: Acquity UPLC BEH C181.7 μm, 50x2.1mm；Eluent A: water+0.1 volume % formic acid (99%), eluent B: acetonitrile；Gradient: 0-1.6min 1-99%B, 1.6-2.0min 99%B；Flow velocity: 0.8mL/min；Temperature: 60 DEG C；Sample size: 2 μ L；DAD scans: 210-400nm；ELSD

Intermediate

Intermediate 1

The bromo-6-of 3-chloro-imidazo [1,2-b] pyridazine

According to described in such as WO 2007/147646 or DE 10 2,006 029447, the following synthesis bromo-6-of 3-is chloro- Imidazo [1,2-b] pyridazine:

The preparation of step 1:6-chlorine imidazo [1,2-b] pyridazine:

By the 2-Chloro-1-ethanal of the 3-amino-6-chlorine pyridazine of 5.0g (38.6mmol) with 4.7mL (40mmol), (in water, 55% contains Amount) in 15mL n-butyl alcohol, jointly heat at 120 DEG C, continue the time of 5 days.After having reacted, reactant mixture is added Saturated sodium bicarbonate solution, and it is extracted with ethyl acetate three times.Then the organic facies saturated nacl aqueous solution merged is washed, And it is dried with sodium sulfate, solvent in vacuo is removed.By, in the chromatographic final purification on silica gel, being separated to 4.17g (70%) the expectation product of amorphous white solid form.

¹H-NMR (chloroform-d): δ [ppm]=7.06 (1H)；7.79(1H)；7.92(1H)；7.96(1H).

The preparation of step 2:3-bromo-6-chlorine imidazo [1,2-b] pyridazine:

In argon, 6-chlorine imidazo [1, the 2-b] pyridazine of 478mg (3.11mmol) is introduced in 10mL chloroform, at ice During middle cooling, add the N-bromosuccinimide of 664mg (3.73mmol).After addition, by reactant mixture in room temperature Under be stirred overnight.Then reactant mixture is mixed with water and ethyl acetate, add saturated sodium bicarbonate solution, then will divide mutually From.Aqueous phase ethyl acetate is extracted three times again.Then the organic facies saturated nacl aqueous solution merged is washed, and use sulfur Acid sodium is dried.Last vacuum removes solvent, separates to obtain the desired product of amorphous white solid form with quantitative yield, will It uses under not further chromatography purification in reaction subsequently.

¹H-NMR (chloroform-d): δ [ppm]=7.12 (1H)；7.79(1H)；7.90(1H).

Intermediate 2

3-(1-benzofuran-2-base)-6-chlorine imidazo [1,2-b] pyridazine

By the bromo-6-of 13.9g (59.8mmol) 3-chloro-imidazo [1,2-b] pyridazine at 508mL 1,4-dioxane Middle suspension.Add 10.1g (62.8mmol) 2-benzofuran ylboronic acid, 2.76g (2.29mmol) tetrakis triphenylphosphine palladium-(0) With 90mL (180mmol) 2M aqueous sodium carbonate.The mixture of gained is heated to 100 DEG C, continues 24 hours.

Add the saturated aqueous ammonium chloride of 400mL.The mixture of gained is extracted with ethyl acetate.Close with saline washing And organic layer and be dried with magnesium sulfate.Solvent is evaporated, then by the solid matter of gained at 40mL dichloromethane and methanol (8:2) mixture impregnates (digest), leaches and be vacuum dried, to obtain the solid matter form of 5.42g (44%) Title compound.

¹H-NMR(300MHz,DMSO-d₆): δ [ppm]=7.23-7.40 (2H), 7.51 (1H), 7.59-7.67 (2H), 7.77(1H),8.33-8.40(2H).

LCMS (method 1): R_t=1.35min；MS (ESIpos) m/z=270 [M+H]⁺.

Intermediate 3

The chloro-3-of 6-(4-methoxyl group-1-benzofuran-2-base) imidazo [1,2-b] pyridazine

It is similar to 3-(1-benzofuran-2-base)-6-chlorine imidazo [1,2-b] pyridazine, by 1.68g (7.22mmol) 3- Bromo-6-chlorine imidazo [1,2-b] pyridazine is prepared the chloro-3-of 6-(4-methoxyl group-1-benzofuran-2-base) imidazo [1,2-b] and is rattled away Piperazine, obtains the solid matter of yield 43%.

¹H-NMR(300MHz,DMSO-d₆), δ [ppm]=3.96 (3H), 6.85-6.91 (1H), 7.25-7.38 (2H), 7.52-7.59(2H),8.37-8.43(2H)

LCMS (method 1): R_t=1.31min；MS (ESIpos) m/z=300 [M+H]⁺.

Intermediate 4

The chloro-3-of 6-(5-methoxyl group-1-benzofuran-2-base) imidazo [1,2-b] pyridazine

It is similar to 3-(1-benzofuran-2-base)-6-chlorine imidazo [1,2-b] pyridazine, bromo-by 1.74g (7.5mmol) 3- 6-chlorine imidazo [1,2-b] pyridazine prepares the chloro-3-of 6-(5-methoxyl group-1-benzofuran-2-base) imidazo [1,2-b] pyridazine, Obtain the solid matter of yield 45%.

¹H-NMR(300MHz,DMSO-d₆), δ [ppm]=3.81 (3H), 6.91-6.99 (1H), 7.33 (1H), 7.50- 7.60(3H),8.35-8.42(2H).

LCMS (method 1): R_t=1.29min；MS (ESIpos) m/z=300 [M+H]⁺.

Intermediate 5

The chloro-3-of 6-(5-chloro-1-benzofuran-2-base) imidazo [1,2-b] pyridazine

Step 1: 2g (13mmol) 7-chloro-1-benzofuran mixture in dry THF (100mL) is cooled to-78 ℃.The n-BuLi of addition 7.9mL (19.7mmol) solution in hexane, and the mixture of gained is stirred at-78 DEG C Continue 1 hour.Add the tributyltin chloride of 5.3mL (19.7mmol).Reactant is stirred at room temperature overnight.

It is carefully added into methanol, and evaporates solvent.The residue obtained is passed through purified by flash chromatography, to obtain 6.2g phase The 2-stannane base benzofuran crude product answered, uses under not being further purified in step 2 by it.

Step 2: in an inert atmosphere, at 85 DEG C, in the manometer tube sealed, by the bromo-6-of 2.34g (10.1mmol) 3- Chloro-imidazo [1,2-b] pyridazine, the crude product 2-stannane base benzofuran of 5.79g (13.1mmol) step 1,192mg (1mmol) Hydro-Giene (Water Science). (I) and 354mg (0.5mmol) double (triphenylphosphine) Palladous chloride. (II) stir in 100mL THF, continue 19h.Steam Send out solvent, the solid obtained extracted in methanol and leaches, obtaining the title compound of 2.73g solid matter form, by it Use in reaction subsequently with the form of crude product.

LCMS (method 3): R_t=1.00min；MS (ESIpos) m/z=304 [M+H]⁺.

Intermediate 6

The chloro-3-of 6-(5-fluoro-1-benzofuran-2-base) imidazo [1,2-b] pyridazine

3-chloro-with 6-(5-chloro-1-benzofuran-2-base) imidazo [1,2-b] pyridazine is similar to, by 513mg (2.21mmol) 3-bromo-6-chlorine imidazo [1,2-b] pyridazine prepares the chloro-3-of 6-(5-fluoro-1-benzofuran-2-base) imidazo [1,2-b] pyridazine, obtains the solid matter (about 57% purity) of 166mg.By this material under not being further purified subsequently Step in use.

LCMS (method 4): R_t=1.37min；MS (ESIpos) m/z=288 [M+H]+.

Intermediate 7

The chloro-3-of 6-(4-fluoro-1-benzofuran-2-base) imidazo [1,2-b] pyridazine

3-chloro-with 6-(5-chloro-1-benzofuran-2-base) imidazo [1,2-b] pyridazine is similar to, by 921mg (3.96mmol) 3-bromo-6-chlorine imidazo [1,2-b] pyridazine prepares the chloro-3-of 6-(4-fluoro-1-benzofuran-2-base) imidazo [1,2-b] pyridazine, obtains the solid matter of 929mg, it is used with the form of crude product.

1H-NMR(300MHz,DMSO-d₆), δ [ppm]=7.09-7.23 (1H), 7.32-7.45 (1H), 7.55 (3H), 8.41(2H).

LCMS (method 3): Rt=1.42min；MS (ESIpos) m/z=288 [M+H]+.

Embodiment

Embodiment 1

3-(1-benzofuran-2-base)-6-[2-(morpholine-2-yl) ethyoxyl] imidazo [1,2-b] pyridazine

In ice bath, 68.1mg (0.52mmol) 2-(2-morpholinyl) ethanol is added in 4mL anhydrous tetrahydro furan 18.3mg (0.46mmol) sodium hydride (in mineral oil 60%) in.Ice bath stirs 15 minutes, is subsequently adding 70mg (0.26mmol) 3-(1-benzofuran-2-base)-6-chlorine imidazo [1,2-b] pyridazine.Ice bath is removed, and by reactant mixture 72h is stirred at 40 DEG C.

Reactant mixture is poured into water, and is extracted with ethyl acetate.Organic layer magnesium sulfate is dried, and concentrates.Will Residue passes through HPLC purification, to obtain the product of the solid matter form of 45mg (47%).

¹H-NMR(400MHz,DMSO-d₆), δ [ppm]=1.86-1.96 (2H), 2.40 (1H), 2.56-2.68 (2H), 2.82(1H),3.43(1H),3.52-3.60(1H),3.73(1H),4.53-4.60(2H),7.01(1H),7.24-7.35 (2H),7.59-7.65(2H),7.67-7.74(1H),8.10-8.18(2H).

LC-MS (method 3): R_t=0.78min；MS (ESIpos) m/z=365 [M+H]⁺.

Embodiment 2

3-(4-methoxyl group-1-benzofuran-2-base)-6-[(2R)-morpholine-2-ylmethoxy] imidazo [1,2-b] is rattled away Piperazine

In ice bath, 191mg (1.6mmol) (R)-2-hydroxymethyl morpholine is added in 24mL anhydrous tetrahydro furan 64mg (1.6mmol) sodium hydride (in mineral oil 60%) in.Ice bath stirs 15 minutes, is subsequently adding 120mg (0.4mmol) the chloro-3-of 6-(4-methoxyl group-1-benzofuran-2-base)-imidazo [1,2-b] pyridazine.Ice bath is removed, and will Reactant mixture is stirred at room temperature 24h.

Reactant mixture is poured in saturated aqueous ammonium chloride, and is extracted with ethyl acetate.By organic layer magnesium sulfate It is dried, and concentrates.By residue by HPLC purification, to obtain the product of the solid matter form of 21mg (14%).

¹H-NMR(300MHz,DMSO-d₆), δ [ppm]=2.63-2.73 (3H), 2.95 (1H), 3.48 (1H), 3.77 (1H),3.92(4H),4.41(2H),6.83(1H),7.04(1H),7.19-7.33(2H),7.53(1H),8.02-8.18 (2H).

LC-MS (method 3): R_t=0.81min；MS (ESIpos) m/z=381 [M+H]⁺.

Embodiment 3

3-(1-benzofuran-2-base)-6-(morpholine-2-ylmethoxy) imidazo [1,2-b] pyridazine

Step 1: in ice bath, adds 2.0g (8.9mmol) 2-(hydroxymethyl) morpholine-4-carboxylic acid tert-butyl ester and extremely exists In 188mg (7.83mmol) sodium hydride (in mineral oil 60%) in 24mL anhydrous tetrahydro furan.15 points are stirred in ice bath Clock, is subsequently adding 1.2g (4.45mmol) 3-(1-benzofuran-2-base)-6-chlorine imidazo [1,2-b] pyridazine.Ice bath is moved Remove, and reactant mixture is stirred at room temperature 4 days.

Reactant mixture is poured in saturated aqueous ammonium chloride, and is extracted with ethyl acetate.The organic facies merged is used Saline washs, and is dried with magnesium sulfate and concentrates.The crude product (3.3g) obtained is made under not being further purified in step 2 With.

Step 2: add 8.9mL trifluoroacetic acid in the crude product of the 2.2g step 1 in 36mL dichloromethane.Will mixing Thing stirring 3h.Add ammonia until mixture reaches alkaline pH.Add saline, and mixture dichloromethane is extracted.To have Machine layer separates, and is dried with magnesium sulfate and concentrates.Obtain the solid matter of 1.68g crude form.

By a small amount of sample (75mg) by HPLC purification, obtain the product of 18mg solid matter form.

¹H-NMR(300MHz,DMSO-d₆), δ [ppm]=2.64-2.75 (3H), 2.94-3.02 (1H), 3.51 (1H), 3.76-3.92(1H),4.45(2H),7.06(1H),7.23-7.37(2H),7.60-7.66(1H),7.72(1H),8.12- 8.19(2H).

LC-MS (method 3): R_t=0.81min；MS (ESIpos) m/z=381 [M+H]⁺.

Embodiment 4

N-(3-{ [3-(1-benzofuran-2-base) imidazo [1,2-b] pyridazine-6-base] epoxide } propyl group)-2,2-diformazan Base propane-1-amine

In ice bath, by 75mg (0.52mmol), (3-[(2,2-dimethyl propyl) amino] propane-1-alcohol adds and extremely exists In 18mg (0.45mmol) sodium hydride (in mineral oil 60%) in 4mL anhydrous tetrahydro furan.Ice bath stirs 15 minutes, It is subsequently adding 70mg (0.26mmol) 3-(1-benzofuran-2-base)-6-chlorine imidazo [1,2-b] pyridazine.Ice bath is removed, and Reactant mixture is stirred at 40 DEG C 16h.

Reactant mixture is poured into water, and is extracted with ethyl acetate.Organic layer magnesium sulfate is dried, and concentrates.Will Residue passes through HPLC purification, to obtain the product of the solid matter form of 56mg (57%).

¹H-NMR(400MHz,DMSO-d₆), δ [ppm]=0.83 (9H), 1.93-2.02 (2H), 2.26 (2H), 2.72 (2H),4.56(2H),7.00(1H),7.24-7.35(2H),7.58(1H),7.60-7.64(1H),7.66-7.70(1H), 8.13(2H).

LC-MS (method 4): R_t=0.90min；MS (ESIpos) m/z=379 [M+H]⁺.

Embodiment 5

3-(5-methoxyl group-1-benzofuran-2-base)-6-[(2R)-morpholine-2-ylmethoxy] imidazo [1,2-b] is rattled away Piperazine

In ice bath, 191mg (1.6mmol) (R)-2-hydroxymethyl morpholine is added in 3mL anhydrous tetrahydro furan 64mg (1.6mmol) sodium hydride (in mineral oil 60%) in.Ice bath stirs 15 minutes, is subsequently adding 120mg (0.40mmol) the chloro-3-of 6-(5-methoxyl group-1-benzofuran-2-base) imidazo [1,2-b] pyridazine.Ice bath is removed, and will Reactant mixture is stirred at room temperature 24h.

Reactant mixture is poured in saturated aqueous ammonium chloride, and is extracted with ethyl acetate.By organic layer magnesium sulfate It is dried, and concentrates.By residue by HPLC purification, to obtain the product of the solid matter form of 20mg (13%).

¹H-NMR(300MHz,DMSO-d₆), δ [ppm]=2.66-2.71 (3H), 2.87-2.96 (1H), 3.41-3.56 (1H),3.79(5H),4.42(2H),6.90(1H),7.04(1H),7.24(1H),7.48-7.58(2H),8.06-8.19 (2H).

LC-MS (method 3): R_t=0.83min；MS (ESIpos) m/z=381 [M+H]⁺.

Embodiment 6

2-{ [3-(1-benzofuran-2-base) imidazo [1,2-b] pyridazine-6-base] epoxide }-N-(Cvclopropvlmethvl) second Amine

In ice bath, 87mg (0.74mmol) 2-[(Cvclopropvlmethvl) amino] ethane-1-alcohol is added to anhydrous at 5mL In 26mg (0.65mmol) sodium hydride (in mineral oil 60%) in oxolane.Ice bath stirs 15 minutes, then adds Enter 100mg (0.37mmol) 3-(1-benzofuran-2-base)-6-chlorine imidazo [1,2-b] pyridazine.Ice bath is removed, and will be anti- Mixture is answered to stir 16h at 40 DEG C.

Reactant mixture is poured in saturated aqueous ammonium chloride, and is extracted with ethyl acetate.By organic layer magnesium sulfate It is dried, and concentrates.By residue by HPLC purification, to obtain the product of the solid matter form of 56mg (43%).

¹H-NMR(300MHz,DMSO-d₆), δ [ppm]=0.08-0.17 (2H), 0.34-0.45 (2H), 0.85-0.98 (1H),2.54(2H),3.11(2H),4.58(2H),7.03(1H),7.23-7.37(2H),7.59-7.66(2H),7.71 (1H),8.12-8.23(2H).

LC-MS (method 4): R_t=0.82min；MS (ESIpos) m/z=349 [M+H]⁺.

Embodiment 7

3-(1-benzofuran-2-base)-6-[(2R)-morpholine-2-ylmethoxy] imidazo [1,2-b] pyridazine

In ice bath, 355mg (2.97mmol) (R)-2-hydroxymethyl morpholine is added in 6mL anhydrous tetrahydro furan 119mg (2.97mmol) sodium hydride (in mineral oil 60%) in.Ice bath stirs 15 minutes, is subsequently adding 200mg (0.74mmol) 3-(1-benzofuran-2-base)-6-chlorine imidazo [1,2-b] pyridazine.Ice bath is removed, and by reactant mixture It is stirred at room temperature 24h.

Reactant mixture is poured in saturated aqueous ammonium chloride, and is extracted with ethyl acetate.By organic layer magnesium sulfate It is dried, and concentrates.By residue by HPLC purification, to obtain the product of the solid matter form of 67mg (25%).

¹H-NMR(300MHz,DMSO-d₆), δ [ppm]=2.62-2.73 (3H), 2.92-3.02 (1H), 3.43-3.57 (1H),3.72-3.93(2H),4.44(2H),7.05(1H),7.21-7.40(2H),7.59-7.66(2H),7.70-7.75 (1H),8.12-8.20(2H).

LC-MS (method 3): R_t=0.78min；MS (ESIpos) m/z=351 [M+H]⁺.

Embodiment 8

3-(1-benzofuran-2-base)-6-{2-[(3R)-morpholine-3-base] ethyoxyl } imidazo [1,2-b] pyridazine

In ice bath, 68mg (0.52mmol) 2-[(3R)-morpholine-3-base] ethanol is added at 4mL anhydrous tetrahydrochysene furan In 18mg (0.45mmol) sodium hydride (in mineral oil 60%) muttered.Ice bath stirs 15 minutes, is subsequently adding 70mg (0.26mmol) 3-(1-benzofuran-2-base)-6-chlorine imidazo [1,2-b] pyridazine.Ice bath is removed, and by reactant mixture 15h is stirred at 40 DEG C.

Reactant mixture is poured into water, and is extracted with ethyl acetate.Organic layer magnesium sulfate is dried, and concentrates.Will Residue passes through purified by flash chromatography, to obtain the product of the solid matter form of 38mg (40%).

¹H-NMR(400MHz,DMSO-d₆), δ [ppm]=1.81 (2H), 2.70-2.78 (2H), 2.85-2.95 (1H), 3.11(1H),3.34-3.38(1H),3.65(1H),3.76(1H),4.59(2H),7.04(1H),7.27-7.38(2H), 7.62-7.68(2H),7.73(1H),8.16(2H).

LC-MS (method 3): R_t=0.73min；MS (ESIpos) m/z=365 [M+H]⁺.

Embodiment 9

3-{ [3-(1-benzofuran-2-base) imidazo [1,2-b] pyridazine-6-base] epoxide }-N-(propane-2-base) third Alkane-1-amine

In ice bath, 89mg (0.74mmol) 3-(propane-2-base amino) propane-1-alcohol is added at 5mL anhydrous four In 26mg (0.65mmol) sodium hydride (in mineral oil 60%) in hydrogen furan.Ice bath stirs 15 minutes, is subsequently adding 100mg (0.37mmol) 3-(1-benzofuran-2-base)-6-chlorine imidazo [1,2-b] pyridazine.Ice bath is removed, and will reaction Mixture is stirred at room temperature 16h.

Reactant mixture is poured in saturated aqueous ammonium chloride, and adds ethyl acetate.The precipitation of gained is leached, uses Water and ethyl acetate washing, and be vacuum dried, to obtain the product of the solid matter form of 124mg (95%).

¹H-NMR(400MHz,DMSO-d₆), δ [ppm]=1.23 (6H), 2.19-2.29 (2H), 3.11 (2H), 4.61 (2H),7.03(1H),7.24-7.36(2H),7.60-7.65(1H),7.67(1H),7.71-7.76(1H),8.14-8.21 (2H).

LC-MS (method 2): R_t=0.85min；MS (ESIpos) m/z=351 [M+H]⁺.

Embodiment 10

N-(2-{ [3-(1-benzofuran-2-base) imidazo [1,2-b] pyridazine-6-base] epoxide } ethyl) propane-2-amine

In ice bath, 78mg (0.74mmol) 2-(isopropylamino) ethane-1-alcohol is added at 5mL anhydrous tetrahydrochysene furan In 26mg (0.65mmol) sodium hydride (in mineral oil 60%) muttered.Ice bath stirs 15 minutes, is subsequently adding 100mg (0.37mmol) 3-(1-benzofuran-2-base)-6-chlorine imidazo [1,2-b] pyridazine.Ice bath is removed, and by reactant mixture It is stirred at room temperature 16h.

Reactant mixture is poured in saturated aqueous ammonium chloride, and is extracted with ethyl acetate.By organic layer magnesium sulfate It is dried, and concentrates.By residue by HPLC purification, to obtain the product of the solid matter form of 65mg (46%).

¹H-NMR(300MHz,DMSO-d₆), δ [ppm]=1.00 (6H), 3.00 (2H), 3.39 (1H), 4.53 (2H), 6.96-7.06(1H),7.23-7.36(2H),7.59-7.66(2H),7.68-7.74(1H),8.12-8.18(2H).

LC-MS (method 4): R_t=0.80min；MS (ESIpos) m/z=337 [M+H]⁺.

Embodiment 11

3-(1-benzofuran-2-base)-6-[(2S)-morpholine-2-ylmethoxy] imidazo [1,2-b] pyridazine

In ice bath, 355mg (2.97mmol) (S)-2-hydroxymethyl morpholine is added in 6mL anhydrous tetrahydro furan 119mg (2.97mmol) sodium hydride (in mineral oil 60%) in.Ice bath stirs 15 minutes, is subsequently adding 200mg (0.74mmol) 3-(1-benzofuran-2-base)-6-chlorine imidazo [1,2-b] pyridazine.Ice bath is removed, and by reactant mixture It is stirred at room temperature 24h.

Reactant mixture is poured in saturated aqueous ammonium chloride, and is extracted with ethyl acetate.By organic layer magnesium sulfate It is dried and concentrates, to obtain the crude product of 280mg.By the crude product of 49mg by HPLC purification, to obtain the solid matter of 2mg The product of form.

¹H-NMR(300MHz,DMSO-d₆), δ [ppm]=2.64-2.72 (3H), 2.94 (1H), 3.42-3.54 (1H), 3.72-3.89(2H),4.44(2H),7.06(1H),7.23-7.36(2H),7.60-7.65(2H),7.73(1H),8.12- 8.20(2H).

LC-MS (method 3): R_t=0.76min；MS (ESIpos) m/z=351 [M+H]⁺.

Embodiment 12

N-(2-{ [3-(1-benzofuran-2-base) imidazo [1,2-b] pyridazine-6-base] epoxide } ethyl)-2,2-diformazan Base propane-1-amine

In ice bath, 68mg (0.52mmol) 2-[(2,2-dimethyl propyl) amino] ethanol is added to anhydrous at 4mL In 18mg (0.46mmol) sodium hydride (in mineral oil 60%) in oxolane.Ice bath stirs 15 minutes, then adds Enter 70mg (0.26mmol) 3-(1-benzofuran-2-base)-6-chlorine imidazo [1,2-b] pyridazine.Ice bath is removed, and will reaction Mixture stirs 72h at 40 DEG C.

Reactant mixture is poured into water, and is extracted with ethyl acetate.Organic layer magnesium sulfate is dried, and concentrates.Will Residue passes through HPLC purification, to obtain the product of the solid matter form of 50mg (53%).

¹H-NMR(300MHz,DMSO-d₆), δ [ppm]=0.84 (9H), 2.35 (2H), 3.01 (2H), 4.55 (2H), 7.03(1H),7.23-7.36(2H),7.62(2H),7.67-7.72(1H),8.11-8.17(2H).

LC-MS (method 4): R_t=0.89min；MS (ESIpos) m/z=365 [M+H]⁺.

Embodiment 13

3-(1-benzofuran-2-base)-6-{2-[(3S)-morpholine-3-base] ethyoxyl } imidazo [1,2-b] pyridazine

In ice bath, 96mg (0.52mmol) (S)-2-(morpholine-3-base) ethanol is added at 4mL anhydrous tetrahydro furan In 18mg (0.46mmol) sodium hydride (in mineral oil 60%) in.Ice bath stirs 15 minutes, is subsequently adding 70mg (0.26mmol) 3-(1-benzofuran-2-base)-6-chlorine imidazo [1,2-b] pyridazine.Ice bath is removed, and by reactant mixture 15h is stirred at 40 DEG C.

Reactant mixture is poured into water, and is extracted with ethyl acetate.Organic layer magnesium sulfate is dried, and concentrates.Will Residue passes through purified by flash chromatography, to obtain the product of the solid matter form of 51mg (54%).

¹H-NMR(400MHz,DMSO-d₆), δ [ppm]=1.81 (2H), 2.73-2.80 (2H), 2.86-2.95 (1H), 3.12(1H),3.33-3.40(1H),3.65(1H),3.76(1H),4.59(2H),7.03(1H),7.27-7.38(2H), 7.62-7.67(2H),7.70-7.75(1H),8.16(2H).

LC-MS (method 3): R_t=0.74min；MS (ESIpos) m/z=365 [M+H]⁺.

Embodiment 14

6-(azetidine-3-ylmethoxy)-3-(1-benzofuran-2-base) imidazo [1,2-b] pyridazine

In ice bath, 64mg (0.52mmol) 3-(hydroxymethyl) azetidine hydrochloride is added to anhydrous at 4mL In 41mg (1.04mmol) sodium hydride (in mineral oil 60%) in oxolane.Ice bath stirs 15 minutes, then adds Enter 70mg (0.26mmol) 3-(1-benzofuran-2-base)-6-chlorine imidazo [1,2-b] pyridazine.Ice bath is removed, and will reaction Mixture stirs 72h at 40 DEG C.

Reactant mixture is poured into water, and is extracted with ethyl acetate.Organic layer magnesium sulfate is dried, and concentrates.Will Residue passes through HPLC purification, to obtain the product of the solid matter form of 39mg (46%).

¹H-NMR(300MHz,DMSO-d₆), δ [ppm]=3.18 (1H), 3.50-3.60 (2H), 3.66-3.77 (2H), 4.63(2H),7.03(1H),7.22-7.37(2H),7.60-7.66(2H),7.70-7.76(1H),8.12-8.19(2H).

LC-MS (method 3): R_t=0.74min；MS (ESIpos) m/z=321 [M+H]⁺.

Embodiment 15

3-(1-benzofuran-2-base)-6-{2-[(2S)-pyrrolidin-2-yl] ethyoxyl } imidazo [1,2-b] pyridazine

Step 1: in 116mL oxolane 9.3g (40.4mmol) [(2S)-1-(tertbutyloxycarbonyl) pyrrolidine- 2-yl] acetic acid dropping 40mL borane-dimethyl sulfide complex.The mixture of gained is stirred at 80 DEG C 2h.

Mixture is poured in saturated sodium bicarbonate aqueous solution carefully.Water layer methyl tertiary butyl ether(MTBE) is extracted.To close And organic layer with saline wash, be dried with magnesium sulfate, and concentrate to obtain the crude product of 6.2g, it be not further purified Under use in step 2.

Step 2: in ice bath, adds the crude product of 1.37g (6.39mmol) step 1 at 34mL anhydrous tetrahydro furan In 224mg (5.62mmol) sodium hydride (in mineral oil 60%) in.Ice bath stirs 15 minutes, is subsequently adding 861mg (3.19mmol) 3-(1-benzofuran-2-base)-6-chlorine imidazo [1,2-b] pyridazine.Ice bath is removed, and by reactant mixture It is stirred at room temperature 24h.

Reactant mixture is poured in saturated aqueous ammonium chloride, and is extracted with ethyl acetate.The organic facies merged is used Saline washs, and is dried with magnesium sulfate and concentrates.The crude product (2.1g) obtained is made under not being further purified in step 3 With.

Step 3: add 4.9mL trifluoroacetic acid in the crude product in the 1.4g step 2 in 28mL dichloromethane.Will be mixed Compound stirring 1h.Add sodium hydrate aqueous solution until mixture reaches alkaline pH.Add saline, and by mixture dichloromethane Alkane extracts.Organic layer is separated, is dried with magnesium sulfate and concentrates.

By residue by HPLC purification, to obtain the product of the solid matter form of 725mg.

1H-NMR(300MHz,DMSO-d₆), δ [ppm]=1.57-1.72 (1H), 1.77-2.01 (2H), 2.11-2.32 (3H),3.09-3.24(2H),3.64(1H),4.51-4.70(2H),7.02(1H),7.24-7.37(2H),7.60-7.66 (2H),7.67-7.74(1H),8.13-8.23(2H).

LC-MS (method 1): R_t=0.82min；MS (ESIpos) m/z=349 [M+H]⁺.

Embodiment 16

3-(1-benzofuran-2-base)-6-(piperidin-2-yl methoxyl group) imidazo [1,2-b] pyridazine

Step 1: in ice bath, adds 1.95g (8.9mmol) 2-(hydroxymethyl) piperidines-1-carboxylic acid tert-butyl ester and extremely exists In 313mg (7.83mmol) sodium hydride (in mineral oil 60%) in 24mL anhydrous tetrahydro furan.15 points are stirred in ice bath Clock, is subsequently adding 1.2g (4.45mmol) 3-(1-benzofuran-2-base)-6-chlorine imidazo [1,2-b] pyridazine.Ice bath is moved Remove, and reactant mixture is stirred at room temperature 4 days.

Reactant mixture is poured in saturated aqueous ammonium chloride, and is extracted with ethyl acetate.The organic facies merged is used Saline washs, and is dried with magnesium sulfate and concentrates.The crude product (1.65g) obtained is made under not being further purified in step 2 With.

Step 2: add 8.9mL trifluoroacetic acid in the crude product of the step 1 in 36mL dichloromethane.Mixture is stirred Mix 3h.Add ammonia until mixture reaches alkaline pH.Add saline, and mixture dichloromethane is extracted.By organic layer Separate, be dried with magnesium sulfate and concentrate.

By residue by HPLC purification, to obtain the product of the solid matter form of 358mg (23%).

¹H-NMR(500MHz,DMSO-d₆), δ [ppm]=1.32-1.49 (3H), 1.62 (1H), 1.84 (2H), 2.66- 2.71(1H),3.09(1H),3.17(1H),4.40-4.45(1H),4.46-4.51(1H),7.07(1H),7.30-7.35 (1H),7.36-7.40(1H),7.65(1H),7.66-7.69(1H),7.74-7.78(1H),8.19-8.23(2H).

LC-MS (method 1): R_t=0.82min；MS (ESIpos) m/z=349 [M+H]⁺.

Embodiment 17

N-(2-{ [3-(1-benzofuran-2-base) imidazo [1,2-b] pyridazine-6-base] epoxide } ethyl) cyclopropylamine

In ice bath, 77mg (0.74mmol) 2-(cyclopropylamino) ethane-1-alcohol is added at 5mL anhydrous tetrahydrochysene furan In 26mg (0.65mmol) sodium hydride (in mineral oil 60%) muttered.Ice bath stirs 15 minutes, is subsequently adding 100mg (0.37mmol) 3-(1-benzofuran-2-base)-6-chlorine imidazo [1,2-b] pyridazine.Ice bath is removed, and by reactant mixture It is stirred at room temperature 16h.

Reactant mixture is poured in saturated aqueous ammonium chloride, and is extracted with ethyl acetate.By organic layer magnesium sulfate It is dried, and concentrates.Residue is extracted in methanol, to obtain the title compound of the solid matter form of 25mg (20%).

¹H-NMR(300MHz,DMSO-d₆), δ [ppm]=0.70-0.94 (4H), 2.82 (1H), 3.58 (2H), 4.81 (2H),7.04(1H),7.24-7.38(2H),7.61-7.67(2H),7.71(1H),8.17-8.25(2H).

LC-MS (method 2): R_t=0.82min；MS (ESIpos) m/z=335 [M+H]⁺.

Embodiment 18

N-(2-{ [3-(1-benzofuran-2-base) imidazo [1,2-b] pyridazine-6-base] epoxide } ethyl)-2-methyl-prop Alkane-2-amine

In ice bath, 61mg (0.52mmol) 2-(tert-butylamino) ethane-1-alcohol is added at 4mL anhydrous tetrahydrochysene furan In 18.3mg (0.46mmol) sodium hydride (in mineral oil 60%) muttered.Ice bath stirs 15 minutes, is subsequently adding 70mg (0.26mmol) 3-(1-benzofuran-2-base)-6-chlorine imidazo [1,2-b] pyridazine.Ice bath is removed, and reaction is mixed Compound stirs 72h at 40 DEG C.

Reactant mixture is poured into water, and is extracted with ethyl acetate.Organic layer magnesium sulfate is dried, and concentrates.Will Residue extracts in methyl tertiary butyl ether(MTBE), to obtain the title compound of the solid matter form of 73mg (80%).

¹H-NMR(400MHz,DMSO-d₆), δ [ppm]=1.05 (9H), 2.96 (2H), 4.49 (2H), 7.02 (1H), 7.24-7.35(2H),7.62(2H),7.68-7.73(1H),8.11-8.16(2H).

LC-MS (method 4): R_t=0.82min；MS (ESIpos) m/z=351 [M+H]⁺.

Embodiment 19

2-{ [3-(1-benzofuran-2-base) imidazo [1,2-b] pyridazine-6-base] epoxide }-N-(propane-2-base) third Alkane-1-amine

In ice bath, 61mg (0.52mmol) 1-(isopropylamino) propane-2-alcohol is added at 4mL anhydrous tetrahydrochysene furan In 18.3mg (0.46mmol) sodium hydride (in mineral oil 60%) muttered.Ice bath stirs 15 minutes, is subsequently adding 70mg (0.26mmol) 3-(1-benzofuran-2-base)-6-chlorine imidazo [1,2-b] pyridazine.Ice bath is removed, and reaction is mixed Compound stirs 16h at 40 DEG C.

Reactant mixture is poured into water, and is extracted with ethyl acetate.Organic layer magnesium sulfate is dried, and concentrates.Will Residue passes through HPLC purification, to obtain the title compound of the solid matter form of 52mg (57%).

¹H-NMR(400MHz,DMSO-d₆), δ [ppm]=0.97 (6H), 1.46 (3H), 2.74-2.84 (2H), 2.96 (1H),5.25-5.35(1H),6.97(1H),7.24-7.35(2H),7.59(1H),7.62(1H),7.71(1H),8.14 (2H).

LC-MS (method 4): R_t=0.84min；MS (ESIpos) m/z=351 [M+H]⁺.

Embodiment 20

3-(5-chloro-1-benzofuran-2-base)-6-[(2R)-morpholine-2-ylmethoxy] imidazo [1,2-b] pyridazine

In ice bath, 189mg (1.58mmol) (R)-2-hydroxymethyl morpholine is added in 4mL anhydrous tetrahydro furan 63mg (1.58mmol) sodium hydride (in mineral oil 60%) in.Ice bath stirs 15 minutes, is subsequently adding 120mg (0.4mmol) the chloro-3-of 6-(5-chloro-1-benzofuran-2-base) imidazo [1,2-b] pyridazine.Ice bath is removed, and reaction is mixed Compound is stirred at room temperature 16h.

Reactant mixture is poured in saturated aqueous ammonium chloride, and is extracted with ethyl acetate.By organic layer magnesium sulfate It is dried, and concentrates.Residue is extracted in methanol, to obtain the title compound of the solid matter form of 15mg (10%).

1H-NMR(300MHz,DMSO-d₆), δ [ppm]=2.57-2.72 (3H), 2.93 (1H), 3.48 (1H), 3.73- 3.88(2H),4.42(2H),7.07(1H),7.33(1H),7.59(1H),7.66(1H),7.80(1H),8.13-8.19(2H).

LC-MS (method 3): R_t=0.88min；MS (ESIpos) m/z=385 [M+H]⁺.

Embodiment 21

3-(1-benzofuran-2-base)-6-[(2R)-pyrrolidin-2-yl methoxyl group] imidazo [1,2-b] pyridazine

In ice bath, 5g (49.4mmol) (R)-2-(hydroxymethyl) pyrrolidine is added at 466mL anhydrous tetrahydrochysene furan In 2.97g (74.2mmol) sodium hydride (in mineral oil 60%) muttered.Ice bath stirs 15 minutes, is subsequently adding 6.67g (24.7mmol) 3-(1-benzofuran-2-base)-6-chlorine imidazo [1,2-b] pyridazine.Ice bath is removed, and will reaction Mixture stirs 16h at 40 DEG C.

Reactant mixture is poured in saline carefully, and is extracted with ethyl acetate.Organic layer sodium sulfate is dried, and Concentrate.By residue by purified by flash chromatography, to obtain the title compound of the solid matter form of 5.6g (68%).

¹H-NMR(300MHz,DMSO-d₆), δ [ppm]=1.46-2.13 (4H), 2.73-2.89 (2H), 3.45-3.57 (1H),4.26-4.33(2H),6.96-7.02(1H),7.29(2H),7.55(1H),7.61(1H),7.69-7.75(1H), 8.12(2H).

LC-MS (method 3): R_t=0.79min；MS (ESIpos) m/z=335 [M+H]⁺.

Embodiment 22

3-(1-benzofuran-2-base)-6-(piperidines-3-base epoxide) imidazo [1,2-b] pyridazine

In ice bath, 5g (49.4mmol) piperidin-3-ol is added to the 2.96g in 500mL anhydrous tetrahydro furan (74.2mmol) in sodium hydride (in mineral oil 60%).Ice bath stirs 15 minutes, is subsequently adding 6.67g (24.7mmol) 3-(1-benzofuran-2-base)-6-chlorine imidazo [1,2-b] pyridazine.Ice bath is removed, and reactant mixture is stirred at 40 DEG C Mix 12h.

Reactant mixture is poured in saline carefully, and is extracted with ethyl acetate.Organic layer sodium sulfate is dried, and Concentrate.Residue is extracted in ethyl acetate, to obtain the title compound of the solid matter form of 5.5g (60%).

¹H-NMR(400MHz,DMSO-d₆), δ [ppm]=1.54-1.83 (3H), 2.23-2.32 (1H), 2.54-2.63 (1H),2.75(1H),2.81-2.89(1H),3.33(2H),5.06(1H),7.00(1H),7.27-7.39(2H),7.54 (1H),7.63-7.67(1H),7.72-7.76(1H),8.13-8.18(2H).

LC-MS (method 3): R_t=0.80min；MS (ESIpos) m/z=335 [M+H]⁺.

Embodiment 23

3-(1-benzofuran-2-base)-6-[(2S)-pyrrolidin-2-yl methoxyl group] imidazo [1,2-b] pyridazine

In ice bath, 5g (49.4mmol) (S)-2-hydroxymethyl-pyrrolidine is added at 466mL anhydrous tetrahydro furan In 2.96g (74.2mmol) sodium hydride (in mineral oil 60%) in.Ice bath stirs 15 minutes, is subsequently adding 6.67g (24.7mmol) 3-(1-benzofuran-2-base)-6-chlorine imidazo [1,2-b] pyridazine.Ice bath is removed, and by reactant mixture 12h is stirred at 40 DEG C.

Reactant mixture is poured in saline carefully, and is extracted with ethyl acetate.Organic layer sodium sulfate is dried, and Concentrate.By residue by purified by flash chromatography, to obtain the title compound of the solid matter form of 6.1g (62%).

¹H NMR(300MHz,DMSO-d₆) δ [ppm]=1.83-2.21 (4H), 3.40-3.56 (2H), 3.58-3.80 (2H),4.17(1H),4.63-5.21(1H),7.03(1H),7.21-7.41(2H),7.49-7.79(3H),7.88-8.07 (2H).

LC-MS (method 3): R_t=0.78min；MS (ESIpos) m/z=335 [M+H]⁺.

Embodiment 24

1-{ [3-(1-benzofuran-2-base) imidazo [1,2-b] pyridazine-6-base] epoxide }-N-methylpropane-2-amine

At 0-5 DEG C, 132mg (1.48mmol) 2-(methylamino) propane-1-alcohol is added in 7.5mL dry DMF In 59.3mg (1.48mmol) sodium hydride (in mineral oil 60%) in.Ice bath stirs 5 minutes, is subsequently adding 200mg (0.74mmol) 3-(1-benzofuran-2-base)-6-chlorine imidazo [1,2-b] pyridazine.Ice bath is removed, and by it at room temperature Stir 1.5 hours.

Reactant mixture is poured in semi-saturation ammonium chloride solution, and be extracted with ethyl acetate four times.Organic by merge Wash with saline, be dried with magnesium sulfate and concentrate.By residue by HPLC purification, to obtain the product of 107.6mg (45%).

¹H-NMR(300MHz,DMSO-d₆), δ [ppm]=1.21 (3H), 2.43 (3H), 3.15-3.28 (1H), 4.46 (2H),7.02(1H),7.23-7.37(2H),7.57-7.65(2H),7.69(1H),8.11-8.20(2H).

LC-MS (method 2): R_t=0.83min；MS (ESIpos) m/z=323 [M+H]⁺.

Embodiment 25

3-(5-chloro-1-benzofuran-2-base)-6-{2-[(2S)-pyrrolidin-2-yl] ethyoxyl } imidazo [1,2-b] Pyridazine

Step 1: in 116mL oxolane 9.3g (40.4mmol) [(2S)-1-(tertbutyloxycarbonyl) pyrrolidine- 2-yl] acetic acid drips 40mL borane-dimethyl sulfide complex.The mixture of gained is stirred at 80 DEG C 2h.

Step 2: in ice bath, adds the crude product of 150mg (0.7mmol) step 1 in 6mL anhydrous tetrahydro furan 37mg (0.93mmol) sodium hydride (in mineral oil 60%) in.Ice bath stirs 15 minutes, is subsequently adding 189mg (0.47mmol) 3-(1-benzofuran-2-base)-6-chlorine imidazo [1,2-b] pyridazine.Ice bath is removed, and by reactant mixture It is stirred at room temperature 18h.

Reactant mixture is poured into water, and is extracted with ethyl acetate.The organic phase with sodium sulfate of merging is dried, and dense Contracting.The crude product (327mg) obtained is used under not being further purified in step 3.

Step 3: add 1.3mL trifluoroacetic acid in the crude product in the 327mg step 2 in 5.8mL dichloromethane.Will Mixture stirring 1.5h.Add ammonia until mixture reaches alkaline pH.Add saline, and mixture dichloromethane is extracted Take.Organic layer is separated, is dried with magnesium sulfate and concentrates.

By residue by HPLC purification, to obtain the product of the solid matter form of 45mg (17%).

¹H-NMR(300MHz,DMSO-d₆), δ [ppm]=1.38-1.53 (1H), 1.67-1.86 (2H), 1.95-2.12 (3H),2.87-3.06(2H),3.31-3.43(2H),4.60(2H),7.02-7.10(1H),7.33-7.41(1H),7.67 (2H),7.79-7.85(1H),8.15-8.23(2H).

LC-MS (method 3): R_t=0.90min；MS (ESIpos) m/z=383 [M+H]⁺.

Embodiment 26

2-{ [3-(1-benzofuran-2-base) imidazo [1,2-b] pyridazine-6-base] epoxide }-N-methylpropane-1-amine

At 0-5 DEG C, 132mg (1.48mmol) 1-(methylamino) propane-2-alcohol is added in 7.5mL dry DMF In 59.3mg (1.48mmol) sodium hydride (in mineral oil 60%) in.Ice bath stirs 5 minutes, is subsequently adding 200mg (0.74mmol) 3-(1-benzofuran-2-base)-6-chlorine imidazo [1,2-b] pyridazine.Ice bath is removed, and by it at room temperature Stir 1.5 hours.

Reactant mixture is poured in semi-saturation ammonium chloride solution, and be extracted with ethyl acetate four times.Organic by merge Wash with saline, be dried with magnesium sulfate and concentrate.By residue by HPLC purification, to obtain the product of 21mg (9%).

¹H-NMR(400MHz,DMSO-d₆), δ [ppm]=1.45 (3H), 2.36 (3H), 2.80-2.87 (1H), 2.90- 2.98(1H),5.33-5.43(1H),6.96(1H),7.24-7.36(2H),7.58(1H),7.60-7.65(1H),7.68- 7.74(1H),8.14(2H).

LC-MS (method 2): R_t=0.83min；MS (ESIpos) m/z=323 [M+H]⁺.

Embodiment 27

Formic acid-N-(2-{ [3-(1-benzofuran-2-base) imidazo [1,2-b] pyridazine-6-base] epoxide }-2-phenyl second Base) propane-2-amine (1:1)

At 0-5 DEG C, 199mg (1.11mmol) 2-(isopropylamino)-1-phenylethanol is added to anhydrous at 7.5mL In 44.5mg (1.11mmol) sodium hydride (in mineral oil 60%) in DMF.Ice bath stirs 5 minutes, is subsequently adding 150mg (0.56mmol) 3-(1-benzofuran-2-base)-6-chlorine imidazo [1,2-b] pyridazine.Ice bath is removed, and by its Stir 3 hours under room temperature.Reactant mixture is poured in semi-saturation ammonium chloride solution, and be extracted with ethyl acetate four times.To close And organic phases washed with brine, be dried with magnesium sulfate and concentrate.By residue by HPLC purification, to obtain 105mg (41%) Product.

¹H-NMR(400MHz,DMSO-d₆), δ [ppm]=1.05 (6H), 2.86-2.97 (1H), 3.00-3.07 (1H), 3.18-3.26(1H),6.11-6.16(1H),7.17(1H),7.25-7.44(6H),7.61(3H),7.75-7.81(2H), 8.11(1H),8.17-8.24(2H).

LC-MS (method 2): R_t=0.98min；MS (ESIpos) m/z=413 [M+H]⁺.

Embodiment 28

N-(2-{ [3-(1-benzofuran-2-base) imidazo [1,2-b] pyridazine-6-base] epoxide }-2-phenylethyl)-2- Methylpropane-1-amine

At 0-5 DEG C, 215mg (1.11mmol) 2-(isobutylamino)-1-phenylethanol is added to anhydrous at 7.5mL In 44.5mg (1.11mmol) sodium hydride (in mineral oil 60%) in DMF.Ice bath stirs 5 minutes, is subsequently adding 150mg (0.56mmol) 3-(1-benzofuran-2-base)-6-chlorine imidazo [1,2-b] pyridazine.Ice bath is removed, and by its Stir 1.5 hours under room temperature.Reactant mixture is poured in semi-saturation ammonium chloride solution, and be extracted with ethyl acetate four times.Will The organic phases washed with brine merged, is dried with magnesium sulfate and concentrates.By residue by HPLC purification, to obtain 101mg (43%) product.

¹H-NMR(400MHz,DMSO-d₆), δ [ppm]=0.84 (6H), 1.61-1.72 (1H), 2.45 (2H), 2.93- 3.00(1H),3.12-3.20(1H),6.09-6.15(1H),7.16(1H),7.24-7.43(6H),7.60(3H),7.74- 7.79(1H),8.10(1H),8.18(1H).

LC-MS (method 2): R_t=1.11min；MS (ESIpos) m/z=427 [M+H]⁺.

Embodiment 29

(-)-2-{ [3-(1-benzofuran-2-base) imidazo [1,2-b] pyridazine-6-base] epoxide }-N-methyl-2-phenyl Ethamine

At 0-5 DEG C, by 56mg (0.371mmol) racemization 2-(methylamino)-1-phenylethanol add to 2.5mL without In 7.4mg (0.185mmol) sodium hydride (in mineral oil 60%) in water DMF.Ice bath stirs 30 minutes, is subsequently adding 50mg (0.185mmol) 3-(1-benzofuran-2-base)-6-chlorine imidazo [1,2-b] pyridazine.Ice bath is removed, and by its Stir 3 hours under room temperature.

Reactant mixture is poured in semi-saturation ammonium chloride solution, and be extracted with ethyl acetate four times.Organic by merge Wash with saline, be dried with magnesium sulfate and concentrate.By residue by HPLC purification, to obtain the product of 44mg (62%).

LC-MS (method 2): R_t=0.99min；MS (ESIpos) m/z=385 [M+H]⁺.

By enantiomer by chirality HPLC (Chiralpak IA 5 μm, 250x30mm, hexane/ethanol 90:10+ 0.1% diethylamine, 40mL/min) separate.

Peak 1:20mg, α=-432.2 ° (1.00；CHCl₃)

¹H-NMR (300MHz, chloroform-d), δ [ppm]=2.55 (3H), 3.04 (1H), 3.28 (1H), 6.16 (1H), 6.90(1H),7.18(1H),7.24-7.35(3H),7.40(2H),7.48-7.57(3H),7.63(1H),7.90(1H),8.09 (1H).

Embodiment 30

N-(2-{ [3-(1-benzofuran-2-base) imidazo [1,2-b] pyridazine-6-base] epoxide }-2-phenylethyl)-2, 2-dimethylpropane-1-amine

At 0-5 DEG C, by 361.6mg (1.48mmol) 2-[(2,2-dimethyl propyl) amino]-1-phenylethanol hydrochloric acid Salt adds to 118.6mg (2.97mmol) sodium hydride (in mineral oil 60%) in 10mL dry DMF.Ice bath stirs Mix 5 minutes, be subsequently adding 200mg (0.74mmol) 3-(1-benzofuran-2-base)-6-chlorine imidazo [1,2-b] pyridazine.By ice Bath removes, and is stirred at room temperature 1.5 hours.Reactant mixture is poured in semi-saturation ammonium chloride solution, and use acetic acid Ethyl ester extracts four times.The organic phases washed with brine that will merge, is dried with magnesium sulfate and concentrates.By residue by HPLC purification, To obtain the product of 186mg (57%).

¹H-NMR(400MHz,DMSO-d₆), δ [ppm]=0.83 (9H), 2.41 (2H), 2.94-3.02 (1H), 3.14- 3.23(1H),6.09-6.16(1H),7.17(1H),7.24-7.44(6H),7.60(3H),7.74-7.79(1H),8.10 (1H),8.19(1H).

LC-MS (method 2): R_t=1.03min；MS (ESIpos) m/z=441 [M+H]⁺.

Embodiment 31

3-(1-benzofuran-2-base)-6-[(3R)-pyrrolidin-3-yl epoxide] imidazo [1,2-b] pyridazine

At 0-5 DEG C, 1.551g (17.80mmol) (3R)-pyrrolidine-3-alcohol is added in 60mL dry DMF In 712mg (17.80mmol) sodium hydride (in mineral oil 60%).Ice bath stirs 5 minutes, is subsequently adding 2.4g (8.90mmol) 3-(1-benzofuran-2-base)-6-chlorine imidazo [1,2-b] pyridazine.Ice bath is removed, and by it at room temperature It is stirred overnight.Reactant mixture is poured in saturated ammonium chloride solution, and extracts ten times with 100mL chloroform.Organic by merge It is dried with magnesium sulfate, and concentrates.Crude product is merged with same amount of second batch under the same conditions.By residue at silicon Purify with dichloromethane and methanol on glue.Concentrate and use 2-isopropoxy propane to extract the flow point collected.Solid is leached, uses Diethyl ether, and be at room temperature vacuum dried, continue a weekend, to obtain the product of 2.55g (43%).

¹H-NMR(400MHz,DMSO-d₆), δ [ppm]=1.90-1.99 (1H), 2.15-2.26 (1H), 2.79-2.88 (1H),2.89-2.98(1H),2.99-3.05(1H),3.22-3.26(1H,and water signal),5.50-5.56 (1H),6.97(1H),7.24-7.35(2H),7.58-7.65(2H),7.74(1H),8.08-8.17(2H).

LC-MS (method 2): R_t=0.81min；MS (ESIpos) m/z=321 [M+H]⁺.

[α]=62.5 °, (methanol, 0.28)

Embodiment 32

3-(1-benzofuran-2-base)-6-[(3R)-piperidines-3-base epoxide] imidazo [1,2-b] pyridazine

At 0-5 DEG C, 200mg (1.45mmol) (3R)-piperidin-3-ol hydrochlorate is added in 7mL dry DMF In 116.3mg (2.91mmol) sodium hydride (in mineral oil 60%).Ice bath stirs 5 minutes, is subsequently adding 196mg (0.73mmol) 3-(1-benzofuran-2-base)-6-chlorine imidazo [1,2-b] pyridazine.Ice bath is removed, and by it at room temperature It is stirred overnight.Reactant mixture is poured in saturated ammonium chloride solution, and is extracted with ethyl acetate four times.The organic facies that will merge Wash twice with saline, be dried with magnesium sulfate and concentrate.By crude product 5mL DMSO process.Solid is leached, and washes with water Wash.It is vacuum dried at 45 DEG C, obtains the product of 155mg (63%).

¹H-NMR (400MHz, chloroform-d), δ [ppm]=1.69 (1H), 1.85-2.03 (2H), 2.22-2.32 (1H), 2.84-2.92(1H),2.92-3.00(1H),3.10(1H),3.35(1H),5.14-5.22(1H),6.80(1H),7.24- 7.36(2H,and chloroform signal),7.45(1H),7.55(1H),7.65(1H),7.90(1H),8.18(1H).

LC-MS (method 2): R_t=0.78min；MS (ESIpos) m/z=335 [M+H]⁺.

Embodiment 33

3-(4-fluoro-1-benzofuran-2-base)-6-[(2R)-morpholine-2-ylmethoxy] imidazo [1,2-b] pyridazine

In ice bath, 51mg (0.43mmol) (2R)-morpholine-2-base methanol is added in 4mL anhydrous tetrahydro furan 17mg (0.43mmol) sodium hydride (in mineral oil 60%) in.Ice bath stirs 15 minutes, is subsequently adding 74mg (0.22mmol) the chloro-3-of 6-(4-fluoro-1-benzofuran-2-base) imidazo [1,2-b] pyridazine.Ice bath is removed, and will reaction Mixture stirs 18h at 40 DEG C.

Reactant mixture is poured into water carefully, and is extracted with ethyl acetate.Organic layer sodium sulfate is dried, and dense Contracting.By residue by HPLC purification, to obtain the title compound of the solid matter form of 49mg (55%).

¹H-NMR(300MHz,DMSO-d₆), δ [ppm]=2.54-2.72 (3H), 2.94 (1H), 3.42-3.54 (1H), 3.77(1H),3.82-3.91(1H),4.43(2H),7.03-7.17(2H),7.35(1H),7.52(1H),7.58(1H), 8.14-8.20(2H).

LC-MS (method 4): R_t=0.81min；MS (ESIpos) m/z=369 [M+H]⁺.

Embodiment 34

3-(5-fluoro-1-benzofuran-2-base)-6-[(2R)-morpholine-2-ylmethoxy] imidazo [1,2-b] pyridazine

In ice bath, 33mg (0.29mmol) (2R)-morpholine-2-base methanol is added in 4mL anhydrous tetrahydro furan 12mg (0.29mmol) sodium hydride (in mineral oil 60%) in.Ice bath stirs 15 minutes, is subsequently adding 69mg (0.14mmol) the chloro-3-of 6-(5-fluoro-1-benzofuran-2-base) imidazo [1,2-b] pyridazine.Ice bath is removed, and will reaction Mixture stirs 18h at 40 DEG C.

Reactant mixture is poured in saturated aqueous ammonium chloride carefully, and is extracted with ethyl acetate.Organic layer is used Sodium sulfate is dried, and concentrates.By residue by HPLC purification, to obtain the title compound of the solid matter form of 14mg (24%) Thing.

¹H-NMR(400MHz,DMSO-d₆), δ [ppm]=2.65-2.77 (3H), 2.99 (1H), 3.47-3.55 (1H), 3.80(1H),3.83-3.91(1H),4.44(2H),7.04-7.10(1H),7.15(1H),7.53(1H),7.60(1H),7.65 (1H),8.14-8.19(2H).

LC-MS (method 4): R_t=0.82min；MS (ESIpos) m/z=369 [M+H]⁺.

Additionally, the compound of the formula (I) of the present invention can be converted into by any means well known by persons skilled in the art Any salt as herein described.Similarly, can by any salt of the compound of the formula (I) of the present invention by those skilled in the art Any means known is converted into free cpds.

The pharmaceutical composition of the compound of the present invention

The invention still further relates to the pharmaceutical composition of the compound comprising one or more present invention.These compositionss can be utilized Desired pharmacotoxicological effect is realized by being administered to patient in need.For purposes of the invention, patient is needs Treat the mammal including people of concrete disease or disease.Therefore, the present invention includes such pharmaceutical composition, its bag Compound or its salt containing the pharmaceutically present invention of acceptable carrier and pharmacy effective dose.Pharmaceutically acceptable carrier is preferred Such carrier, its under the concentration consistent with the effective active of active component to patient's relative nontoxic and harmless so that Any side effect caused by described carrier will not destroy the beneficial effect of described active component.The pharmacy effective dose of compound is excellent Choosing is the concrete patient's condition treated to produce result or produces the amount of impact.Can use and include that rapid release, slow release and timing are released Put preparation at the most effective interior conventional dosage unit forms, the compound of the present invention pharmaceutically may be used with well known in the art The carrier accepted is administered the most as follows: oral, parenteral, locally, nasal cavity, eye (ophthalmically), eye (optically), Sublingual, rectum, vagina administration etc..

For oral administration, described compound can be configured to solid or liquid preparation, such as capsule, pill, tablet, Containing lozenge (troche), lozenge (lozenge), melten gel agent (melt), powder, solution, suspensoid or Emulsion, and can basis Prepared by the method for preparing pharmaceutical composition known in the art.Solid unit dosage form can be capsule, and it can be common Ebonite bladder type or soft-capsule type, comprise such as surfactant, lubricant and inert filler (such as lactose, sucrose, phosphoric acid Calcium and corn starch).

In another embodiment, can be by the compound of the present invention and conventional tablet bases (such as lactose, sucrose and Semen Maydis Starch) together and it is pressed into tablet with following combinations of substances: binding agent (such as arabic gum, corn starch or gelatin), it is used for The decomposition of tablet and disintegrating agent (such as potato starch, alginic acid, corn starch and guar gum, the Radix astragali tree of dissolution after adjunctive administration Glue, arabic gum), for improving the mobility of tablet granulation and preventing the surface of tablet material and tablet mould and drift from gluing Attached lubricant (such as Talcum, stearic acid or magnesium stearate, calcium stearate or zinc stearate), dyestuff, coloring agent, and be used for (such as Oleum menthae, wintergreen oil or Fructus Pruni pseudocerasi are fragrant for the organoleptic properties improving tablet the flavoring agent making them be easier to be accepted by patients Essence).The excipient being suitable for for oral liquid dosage forms includes dicalcium phosphate and diluent, such as water and alcohol (such as ethanol, benzene Methanol and polyvinyl alcohol), add or without pharmaceutically acceptable surfactant, suspending agent or emulsifying agent.Can exist Other materials various are as coating or for changing the physical form of dosage unit.Such as can use Lac, sugar or the two is by sheet Agent, pill or capsule coating.

Dispersible powder and granule are suitable for preparing aqueous suspension.They provide with dispersant or wetting agent, help Suspension and the active component of one or more preservative mixing.The dispersant being suitable for or the example of wetting agent and suspending agent are upper Literary composition mention those.Also can there is other excipient, such as those described above sweeting agent, flavoring agent and coloring agent.

The pharmaceutical composition of the present invention can be also the form of oil in water emulsion.Oil phase can be vegetable oil, such as liquid paraffin Or the mixture of vegetable oil.The emulsifying agent being suitable for can be (1) natural gum, and such as Radix Acaciae senegalis and gum tragacanth, (2) are natural Phospholipid, such as soybean phospholipid and lecithin, (3) are derived from fatty acid and the ester of hexitan or partial ester, such as Sorbitan The condensation product of alcohol monoleate, (4) described partial ester and oxirane, such as polyoxyethylene sorbitan monooleate.Institute State Emulsion and also can comprise sweeting agent and flavoring agent.

Can be by described active component being suspended in vegetable oil (such as Oleum Arachidis hypogaeae semen, olive oil, Oleum sesami or Oleum Cocois) Or it is suspended in mineral oil (such as liquid paraffin) and prepares Oil suspensions.Described Oil suspensions can comprise thickening agent, Such as Cera Flava, hard paraffin or spermol.Described suspensoid also can comprise one or more preservative, such as P-hydroxybenzoic acid second Ester or P-hydroxybenzoic acid n-propyl；One or more coloring agent；One or more flavoring agents；And one or more sweet tastes Agent, such as sucrose or saccharin.

Available sweeting agent (such as glycerol, propylene glycol, Sorbitol or sucrose) comes syrup blend agent and elixir.This type of preparation Also can comprise demulcent and preservative (such as methyl hydroxybenzoate and propylparaben) and flavoring agent and coloring agent.

Also the compound of the present invention can be carried out parenteral, the most subcutaneous, vein with the injection dosage of described compound In, ophthalmic, intrasynovial, intramuscular or Intraperitoneal medication, described injection dosage preferably can accept the physiology containing pharmaceutical carrier Diluent in, described pharmaceutical carrier can be the mixture of sterile liquid or liquid, described liquid such as water, saline, glucose Aqueous solution and relevant sugar juice, alcohol such as ethanol, isopropanol or hexadecanol, glycol such as propylene glycol or Polyethylene Glycol, glycerol Ketal such as 2,2-dimethyl-1,1-dioxolanes-4-methanol, ether such as PEG 400, oil, fatty acid, fatty acid ester Or fatty glyceride or acetylated fatty acid glyceride, described diluent adds or without there being pharmaceutically acceptable surface Activating agent, such as soap or detergent, suspending agent such as pectin, carbomer, methylcellulose, hypromellose or carboxymethyl Cellulose, or emulsifying agent and other pharmaceutical auxiliaries.

Can be used for the exemplary oil in the parenteral administration of the present invention is that those come from oil, animal, plant or synthesis The oil in source, such as Oleum Arachidis hypogaeae semen, soybean oil, Oleum sesami, Oleum Gossypii semen, Semen Maydis oil, olive oil, vaseline oil and mineral oil.It is suitable for Fatty acid include oleic acid, stearic acid, isostearic acid and myristic acid.The fatty acid ester being suitable for is such as ethyl oleate and lima bean Cool isopropyl propionate.The soap being suitable for includes fatty acid alkali metal salt, ammonium salt and triethanolamine salt, and the detergent being suitable for includes Cationic detegent, such as dimethyl dialkyl ammonium halide, alkylpyridinium halides and alkylamine acetate；Anionic detergent Such as alkylsulfonate, arylsulphonate and alkene sulfonate, alkyl sulfate and alkyl sulfo succinate, olefin sulphates With alkene sulfosuccinate, ether sulfate and sulfosuccinates and monoglyceride sulfates and monoglyceride sulphur Base succinate；Non-ionic detergent, such as fatty amine oxide, fatty acid alkanol amides and poly-(oxygen ethylene-oxygen third Alkene), ethylene oxide copolymer or epoxy propane copolymer；And ampholytic detergent, such as alkyl-Beta-alanine salt and 2-alkane Base imidazoline quaternary ammonium salt, and its mixture.

The parenteral composition of the present invention would generally comprise the described work of about 0.5 weight %-about 25 weight % in the solution Property composition.Preservative and buffer agent are also advantageously used.In order to minimize or eliminate the stimulation to injection site, this type of combination Thing can comprise hydrophil lipophil balance (HLB) and be preferably from about the nonionic surfactant of 12-about 17.In this type of preparation, live in surface The amount of property agent is preferably from about 5 weight %-about 15 weight %.Described surfactant can be the one-component with above HLB, or Person has the mixture of component of desired HLB for two or more.

Exemplary surfactants for parenteral administration is polyethylene sorbitan fatty acid ester, the most de- Water sorbitol monooleate, and the high molecular weight adducts of oxirane and hydrophobic base, described hydrophobic base by Expoxy propane and propylene glycol condensation are formed.

Described pharmaceutical composition can be the form of Injectable sterile aqueous suspension.Can use as follows according to known method Material prepares this type of suspensoid: applicable dispersant or wetting agent and suspending agent, such as sodium carboxymethyl cellulose, Methyl cellulose Element, hypromellose, sodium alginate, polyvinylpyrrolidone, gum tragacanth and Radix Acaciae senegalis；Dispersant or wetting agent, its Can be natural phospholipid (such as lecithin), oxyalkylene and the condensation product (such as Myrj 45) of fatty acid, epoxy Ethane and the condensation product (such as heptadecaethylene oxycetanol) of long-chain fatty alcohol, oxirane with derived from fatty acid and oneself The condensation product (such as polyoxyethylene 80 sorbitan monooleate) of the partial ester of sugar alcohol or oxirane with derived from fatty acid and The condensation product (such as Polysorbate 80) of the partial ester of hexitan.

Aseptic injection preparation can be also that the Injectable sterile in the nontoxic acceptable diluent of parenteral or solvent is molten Liquor or suspensoid.Spendable diluent and solvent are such as water, Ringer's solution, isotonic sodium chlorrde solution and isotonic glucose Solution.It addition, aseptic expressed oi is routinely used for solvent or suspension media.Thus, any zest can be used little Expressed oi, including synthesis monoglyceride or diglyceride.It addition, can be by fatty acid (such as oleic acid) for injection Preparation in.

Also the compositions of the present invention can be administered for the form of the suppository of the rectally of medicine.Can be by by medicine With at normal temperatures for solid but for liquid and therefore can melt in the rectum and discharge described medicine under rectal temperature These compositionss are prepared in the non-irritating excipient mixing being suitable for.This type of material is such as cocoa butter and Polyethylene Glycol.

The another kind of preparation used in the method for the present invention utilizes transdermal delivery device (" patch ").This type of transdermal patch can For providing the continuously or discontinuously infusion of the compound of the present invention of controlled amounts.For delivering the structure of the transdermal patch of medicament With purposes be well known in the art (see for example on June 11st, 1991 bulletin the 5th, 023, No. 252 United States Patent (USP), helped Draw addition herein).This type of patch can be configured for continuously, pulsed or the medicament of on-demand delivery.

Controlled release preparation for parenteral includes liposome microsphere known in the art, polymer microballoon and polymer Gel preparation.

May need maybe by mechanical delivery device, described pharmaceutical composition must be delivered to patient.For delivering medicament The structure of mechanical delivery device and purposes be well known in the art.The direct technology that medicine is such as administered directly to brain is usual Relate to the ventricular system that drug delivery tube is inserted patient to walk around blood brain barrier.For by specific to health of medicament transport This type of implanted delivery system a kind of of anatomical location is recorded in No. 5,011,472 U.S. of bulletin on April 30th, 1991 Patent.

The compositions of the present invention or the most also must can comprise other routine of commonly referred to as carrier or diluent Pharmaceutically acceptable formulation ingredients.The routine operation of dosage form being prepared as such composition being suitable for can be used.Specific examples of such components Including, with operation, those that be recorded in following list of references, described list of references all quotes addition herein: Powell, M.F. etc. People, " Compendium of Excipients for Parenteral Formulations " PDA Journal of Pharmaceutical Science&Technology 1998,52(5),238-311；Strickley,R.G"Parenteral Formulations of Small Molecule Therapeutics Marketed in the United States (1999)-Part-1"PDA Journal of Pharmaceutical Science&Technology 1999,53(6), 324-349；And Nema, S. et al., " Excipients and Their Use in Injectable Products " PDA Journal of Pharmaceutical Science&Technology 1997,51(4),166-171。

Can be used for, time suitably, the common drug composition described compositions being configured to for intended route of administration to include:

Acidulant (example includes but not limited to acetic acid, citric acid, fumaric acid, hydrochloric acid, nitric acid)；

Basifier (example include but not limited to ammonia, ammonium carbonate, diethanolamine, monoethanolamine, potassium hydroxide, sodium borate, Sodium carbonate, sodium hydroxide, triethanolamine (triethanolamine), triethanolamine (trolamine))；

Adsorbent (example includes but not limited to Powderd cellulose and activated carbon)；

(example includes but not limited to carbon dioxide, CCl to aerosol propellant₂F₂、F₂ClC-CClF₂And CClF₃)；

Drive air agent (air displacement agent) (example includes but not limited to nitrogen and argon)；

Antifungal preservative (example include but not limited to benzoic acid, butoben, ethyl hydroxybenzoate, methyl hydroxybenzoate, Propylparaben, sodium benzoate)；

(example includes but not limited to the tertiary fourth of benzalkonium chloride, benzethonium chloride, benzyl alcohol, cetylpyridinium chloride, trichlorine to antibiotic antiseptic Alcohol, phenol, phenethanol, phenylmercuric nitrate and thimerosal)；

Antioxidant (example include but not limited to ascorbic acid, ascorbyl palmitate, butylated hydroxyanisole, butylated hydroxytoluene, Hypophosphorous acid, thioglycerol, propylgallate, sodium ascorbate, sodium sulfite, sodium sulfoxylate formaldehyde, sodium pyrosulfite)；

Adhesion substance (example include but not limited to block polymer, natural and synthetic rubber, polyacrylate, polyurethane, Silicone, polysiloxanes and SB)；

(example includes but not limited to potassium metaphosphate, dipotassium hydrogen phosphate, sodium acetate, anhydrous citric acid sodium and lemon to buffer agent Lemon acid sodium dihydrate)；

(example includes but not limited to syrup acacia, aromatic syrup, aromatic elixir, cherry syrup, cocoa to carrier Syrup, orange syrup, syrup, Semen Maydis oil, mineral oil, Oleum Arachidis hypogaeae semen, Oleum sesami, antibacterial sodium chloride injection and antibacterial injection With water)；

Chelating agen (example includes but not limited to disodium edetate and edetic acid)；

Coloring agent (example include but not limited to FD&C Red No.3, FD&C Red No.20, FD&C Yellow No.6, FD&C Blue No.2, D&C Green No.5, D&C Orange No.5, D&C Red No.8, caramel and iron oxide red)；

Clarifier (example includes but not limited to bentonite)；

(example includes but not limited to arabic gum, cetomacrogol, spermol, glyceryl monostearate, ovum phosphorus to emulsifying agent Fat, dehydrated sorbitol mono-fatty acid ester, polyoxyethylene 50 monostearate)；

Become capsule (example includes but not limited to gelatin and cellulose acetate-phthalate)；

Spice (example includes but not limited to Oleum Anisi Stellati, Oleum Cinnamomi, cocoa, menthol, orange oil, Oleum menthae and vanillin)；

Wetting agent (example includes but not limited to glycerol, propylene glycol and Sorbitol)；

Grinding agent (example includes but not limited to mineral oil and glycerol)；

(example includes but not limited to Oleum Arachidis hypogaeae semen (arachis oil), mineral oil, olive oil, Oleum Arachidis hypogaeae semen (peanut to oil Oil), Oleum sesami and vegetable oil)；

(example includes but not limited to lanoline, hydrophilic ointment, polyethylene glycol ointment, vaseline oil, hydrophilic all to ointment base Intellectual circle's oil, simple ointment, yellow ointment and cold cream)；

(example includes but not limited to monohydroxy or polyhydroxy alcohols, monovalence or multivalence alcohol to penetration enhancers (transdermal delivery) Class, saturated or unsaturated fatty acids alcohols, saturated or unsaturated fatty acids esters, saturated or unsaturated dicarboxylic class, quintessence oil class, phospholipid Acyl derivative, cephalin, terpenoid, amide-type, ethers, ketone and ureas)；

Plasticizer (example includes but not limited to diethyl phthalate and glycerol)；

(example includes but not limited to ethanol, Semen Maydis oil, Oleum Gossypii semen, glycerol, isopropanol, mineral oil, oleic acid, Semen arachidis hypogaeae to solvent Oil, purified water, water for injection, sterile water for injection and Sterile Water for Irrigation)；

Sclerosing agent (example include but not limited to spermol, cetyl esters wax, microwax, paraffin, stearyl alcohol, white beeswax and Cera Flava)；

Suppository base (example includes but not limited to cocoa butter and Polyethylene Glycol (mixture))；

Surfactant (example include but not limited to benzalkonium chloride, nonoxinol 10, octoxinol (oxtoxynol) 9, Polyoxyethylene sorbitan monoleate, sodium lauryl sulphate and span 40)；

(example includes but not limited to agar, bentonite, carbomer, sodium carboxymethyl cellulose, hydroxy ethyl fiber to suspending agent Element, hydroxypropyl cellulose, hypromellose, Kaolin, methylcellulose, Tragacanth and aluminium-magnesium silicate)；

(example includes but not limited to aspartame, glucose, glycerol, mannitol, propylene glycol, saccharin sodium, Pyrusussuriensis to sweeting agent Sugar alcohol and sucrose)；

Tablet antitack agent (example includes but not limited to magnesium stearate and Talcum)；

(example includes but not limited to arabic gum, alginic acid, sodium carboxymethyl cellulose, sompressible sugar, ethyl to tablet binder Cellulose, gelatin, liquid glucose, methylcellulose, non-crosslinked polyvinylpyrrolidone and pregelatinized Starch)；

(example includes but not limited to that calcium hydrogen phosphate, Kaolin, lactose, mannitol, crystallite are fine for tablet and capsule diluent Dimension element, Powderd cellulose, winnofil, sodium carbonate, sodium phosphate, Sorbitol and starch)；

(example includes but not limited to liquid glucose, hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxypropyl in tablet coating agent Methylcellulose, methylcellulose, ethyl cellulose, cellulose acetate-phthalate and Lac)；

Tablet direct pressing excipient (example includes but not limited to calcium hydrogen phosphate)；

(example includes but not limited to alginic acid, carboxymethylcellulose calcium, microcrystalline Cellulose, Po Lakelin potassium to tablet disintegrant (polacrillin potassium), crospolyvinylpyrrolidone, sodium alginate, primojel and starch)；

Tablet fluidizer (example includes but not limited to silica sol, corn starch and Talcum)；

(example includes but not limited to calcium stearate, magnesium stearate, mineral oil, stearic acid and stearic acid to tablet lubricants Zinc)；

Tablets/capsules agent opacifier (example includes but not limited to titanium dioxide)；

Tablet polishing agent (example includes but not limited to Brazil wax and white beeswax)；

Thickening agent (example includes but not limited to Cera Flava, spermol and paraffin)；

Tonicity agent (example includes but not limited to glucose and sodium chloride)；

Viscosifier (example include but not limited to alginic acid, bentonite, carbomer, sodium carboxymethyl cellulose, methylcellulose, Polyvinylpyrrolidone, sodium alginate and Tragacanth)；And

(example includes but not limited to heptadecaethylene oxycetanol (heptadecaethylene to wetting agent Oxycetanol), lecithin, sorbitol monooleate, polyoxyethylene 80 sorbitan monooleate and polyoxyethylene 8 stearate Ester).

The pharmaceutical composition of the present invention can be exemplified below:

Aseptic intravenous solution agent: can use sterile water for injection prepare the present invention expectation compound 5mg/mL molten Liquid, can optionally regulate pH.With aseptic 5% glucose, described solution is diluted to 1-2mg/mL be used for being administered, and about 60 In minute, the form with intravenous infusion is administered.

Lyophilized powder for intravenous administration: the expectation of the present invention of the lyophilized powder form of available (i) 100-1000mg Compound, (ii) 32-327mg/mL sodium citrate, and (iii) 300-3000mg Gentran 40 prepare sterile preparation.Use aseptic injection Concentration said preparation redissolved to 10-20mg/mL with saline or 5% glucose, then the dilutest with saline or 5% glucose Release to 0.2-0.4mg/mL, and intravenous push or administered by infusion in 15-60 minute angular vein.

Intramuscular injection suspensoid: following solution or suspensoid can be prepared for intramuscular injection:

The compound of the desired water-insoluble present invention of 50mg/mL

5mg/mL sodium carboxymethyl cellulose

4mg/mL TWEEN 80

9mg/mL sodium chloride

9mg/mL benzyl alcohol

Hard capsule: by each personal 100mg divided active component, 150mg lactose, 50mg cellulose and 6mg stearic acid Magnesium is filled the two-piece type hard capsule of standard and is prepared substantial amounts of unit capsules.

Soft capsule: prepare active component mixing in digestible oil (such as soybean oil, Oleum Gossypii semen or olive oil) Thing and comprise the soft capsule of active component described in 100mg to be formed in injected the gelatin of fusing by positive displacement pump.By capsule Wash and be dried.Described active component can be dissolved in the mixture of Polyethylene Glycol, glycerol and Sorbitol and mix preparing water Soluble drug mixture.

Tablet: prepare a large amount of tablet by routine operation so that dosage unit comprises 100mg active component, 0.2mg colloid Silicon dioxide, 5mg magnesium stearate, 275mg microcrystalline Cellulose, 11mg starch and 98.8mg lactose.Can use suitable aqueous and Non-aqueous coatings is to increase palatability, improve outward appearance and stability or postpone to absorb.

Quick-release tablet/capsule: these are the solid oral dosage forms prepared by conventional method and new method.It is not required to use water And by these unit oral, for dissolution at once and the delivery of medicine.Described active component is blended in comprise the most sugared, bright In the liquid of the composition of glue, pectin and sweeting agent.These liquid curings are made to become solid by lyophilization with solid state extraction techniques Tablet or caplet.Can be by tabletting together with sugared with viscoelasticity and thermoelastic for medical compounds and polymer or effervescence component with preparation The porous matrix of rapid release under conditions of need not water.

Combined therapy

Term in the present invention " combines " and uses as known in the art, and can be with fixed Combination, on-fixed Presented in combination or medicine box.

" fixed Combination " in the present invention uses as known in the art, and it is alive to be defined as wherein said first Property composition and described second active component are collectively reside in the combination in a unit dosage forms or single entities." fixed Combination " One example is pharmaceutical composition, and wherein said first active component and described second active component are present in for being administered simultaneously Mixture, the most in the formulation.Another example of " fixed Combination " is drug regimen, wherein said first active component and institute State the second active component to be present in a unit, but be not the form of mixture.

Non-fixed combinations or " medicine box " in the present invention use as known in the art, and are defined as wherein institute State the first active component and described second active component is present in more than one unit.Non-fixed combinations or a reality of medicine box Example is combination, and wherein said first active component and described second active component are independently present.Non-fixed combinations or medicine box Component can (chronologically individually, sequentially, simultaneously, concurrently or the most alternately Staggered) it is administered.

The compound of the present invention can be administered as single medicament or be administered with one or more other pharmaceutical agent combinations, its Described in combination will not cause unacceptable untoward reaction.The invention still further relates to this type of combination.Such as, can be by the change of the present invention Compound and known chemotherapeutics or anticarcinogen (the most anti-excess proliferative disease or the medicament etc. of other indication) and and it Mixture and combination be combined.Other indication medicament includes but not limited to that anti-angiogenic agent, mitosis suppress Agent, alkylating agent, antimetabolite, DNA-embed antibiotic, growth factor receptor inhibitors, cell cycle inhibitor, enzyme inhibitor, topology Isomerase inhibitors, biological response modifier or hormone antagonist.

Term " chemotherapeutic anti-cancer agent " includes but not limited to: 131I-chTNT, 1: PN: WO02056903 PAGE: 25 claimed protein, abiraterone, aclarubicin, Ah Ground interleukin, A Lun pearl monoclonal antibody, alitretinoin, altretamine, aminoglutethimide, amrubicin, amsacrine, Anastrozole, Arglabin, arsenic trioxide, asparaginase, azacitidine, basiliximab, BAY 80-6946, BAY 1000394, BAY 86-9766 (RDEA 119, Belotecan, bendamustine, Avastin, bexarotene, bicalutamide, bisantrene, Bleomycin, bortezomib, buserelin, busulfan, Cabazitaxel, calcium folinate, calcium levofolinate, capecitabine, carboplatin, Carmofur, carmustine, card appropriate rope monoclonal antibody, celecoxib, celmoleukin, Cetuximab, chlorambucil, chlorine ground is pregnant Ketone, chlormethine, cisplatin, carat found shore, clodronic acid pamidronic acid, clofarabine, crisantaspase, cyclophosphamide, the special dragon of ring third, arabinose Cytidine, dacarbazine, actinomycin D, reach erythropoietin α, Dasatinib, daunorubicin, decitabine, Ac-D-2Nal-D-4Cpa-D-3Pal-Ser-4Aph(Hor)-D-4Aph(Cbm)-Leu-Lys(iPr)-Pro-D-Ala-NH2, Buddhist nun white Interleukin 2, Shu Dankang, deslorelin, dibrospidium chloride, docetaxel, doxifluridine, doxorubicin, doxorubicin+female Ketone, according to storehouse pearl monoclonal antibody, male according to bending Lip river monoclonal antibody, elliptinium acetate, Ai Qu pool handkerchief, endostatin, enocitabine, epirubicin, epithio Alcohol, erythropoietin α, epoetin beta, Ai Bo, Ai Libulin, Erlotinib, estradiol, estramustine, etoposide, Yi Weimo Department, exemestane, fadrozole, filgrastim, fludarabine, fluorouracil, flutamide, formestane, fotemustine, fluorine dimension department Group, Ganite (Fujisawa)., ganirelix, gefitinib, gemcitabine, lucky trastuzumab, glutoxim, goserelin, Maxamine, Histrelin, hydroxyurea, I-125 seed (I-125seeds), ibandronic acid, ibritumomab tiuxetan, idarubicin, different ring phosphinylidyne Amine, imatinib, miaow quinoline not moral, an improsulfan, interferon-ALPHA, interferon beta, interferon gamma, her wooden monoclonal antibody, irinotecan, she Husky grand, Lanreotide, Lapatinib, lenalidomide, lenograstim, lentinan, letrozole, leuprorelin, levamisole, profit Relax urea, lobaplatin, lomustine, lonidamine, masoprocol, medroxyprogesterone, megestrol, melphalan, mepitiostane, mercaptopurine, Methotrexate, methoxsalen, methylamino ketone valerate, methyltestosterone, meter Fa Mo peptide, miltefosine, rice stand platinum, mitobronitol, Mitoguazone, mitolactol, mitomycin, mitotane, mitoxantrone, nedaplatin, nelarabine 506u, AMN107, nilutamide, Buddhist nun's trastuzumab, nimustine, nitracrine, method wood monoclonal antibody, omeprazole, oprelvekin, oxaliplatin, p53 base difficult to understand Because for the treatment of, paclitaxel, Pa Lifuming, Pd-103 seed (palladium-103seed), Pamidronic Acid, handkerchief wood monoclonal antibody, handkerchief azoles handkerchief Buddhist nun, pegaspargase, PEG-epoetin beta (methoxyl group PEG-epoetin beta), Pei Feisi booth, training Interferon Alpha-2b, the U.S. song of training Plug, pentazocine, pentostatin, peplomycin, perfosfamide, Picibanil, pirarubicin, Plerixafor, plicamycin, poly- Ammonia glucose, polyestradiol phosphate, polysaccharide-k, porfimer sodium, Pralatrexate, prednimustine, procarbazine, quinoline Gao Lai, Lei Luo Former times sweet smell, Raltitrexed, Ranimustine, razoxane, Rui Gefeini, risedronic acid, Rituximab, sieve meter are new, Luo meter Si booth, Sargramostim, sipuleucel-T, sizofiran, sobuzoxane, CMNa, Sorafenib, streptozocin, Sutent, he Draw pool sweet smell, Tamibarotene, tamoxifen, tasonermin, teceleukin, ftorafur, ftorafur+gimeracil+Austria for drawing West, temoporfin, temozolomide, CCI-779, teniposide, testosterone, tetrofosmin, thalidomide, phosphinothioylidynetrisaziridine, thymalfasin, Thioguanine, torr pearl monoclonal antibody, hycamtin, toremifene, tositumomab, ET-743, Herceptin, treosulfan, Retinoic acid, trilostane, triptorelin, trofosfamide, tryptophan, ubenimex, valrubicin, Fan Tanibu, vapreotide, Vemurafenib, vinblastine, vincristine, vindesine, vinflunine, vinorelbine, SAHA, R 83842,90Y Glass microsphere, zinostatin, zinostatin ester, zoledronic acid, zorubicin.

Also can be by the compound of the present invention and protein therapeutic agent administering drug combinations.It is applicable to treat cancer or other blood vessel is raw This type of protein therapeutic agent that one-tenth disease and the compositions with the present invention that is suitable to are used together includes but not limited to interferon (example Such as interferon-alpha, interferon-β or IFN-γ), super excitability (supraagonistic) monoclonal antibody, Tuebingen, TRP-1 protein vaccine, Colostrinin, anti-FAP antibody, YH-16, lucky trastuzumab, infliximab, list of western appropriate former times Anti-, Herceptin, denileukin diftitox, Rituximab, α 1 thymosin, Avastin, mecasermin, woods mecasermin (mecasermin rinfabate), oprelvekin, natalizumab, rhMBL, MFE-CP1+ZD-2767-P, ABT- 828, ErbB2-specific immune toxin, SGN-35, MT-103, Lin Feipei (rinfabate), AS-1402, B43-dyewood Huang Ketone, the radioimmunotherapy agent of L-19 system, AC-9301, NY-ESO-1 vaccine, IMC-1C11, CT-322, rhCC10, r (m) CRP, MORAb-009, A Weikuming, MDX-1307, Her-2 vaccine, APC-8024, NGR-hTNF, rhH1.3, IGN-311, endothelium press down Element, volt Lip river former times monoclonal antibody, PRO-1762, next husky wood monoclonal antibody, SGN-40, pertuzumab, EMD-273063, L19-IL-2 merge egg In vain, PRX-321, CNTO-328, MDX-214, for add pool peptide, CAT-3888, draw shellfish pearl monoclonal antibody, launch a particle radioactivity with The lintuzumab of position element crosslinking, EM-1421, HyperAcute vaccine, celmoleukin monoclonal antibody, galiximab, HPV-16- E7, Javelin-carcinoma of prostate, Javelin-melanoma, NY-ESO-1 vaccine, EGF vaccine, CYT-004-MelQbG10, WT1 Peptide, Ao Gefu monoclonal antibody, method wood monoclonal antibody difficult to understand, prick calamite monoclonal antibody, the pungent interleukin of shellfish, WX-G250, Albuferon, VEGF Trap, Shu Dankang, vaccine, CTP-37, Yi Fengu monoclonal antibody or 131I-chTNT-1/B.Monoclonal antibody bag as protein therapeutic agent Include but be not limited to muromonab-CD3, abciximab, according to bend Lip river monoclonal antibody, daclizumab, gemtuzumab Ozogamicin Mylotarg CDP 771 (gentuzumab), Ah Logical sequence pearl monoclonal antibody, ibritumomab tiuxetan (ibritumomab), Cetuximab, Avastin (bevicizumab), in accordance with the law pearl list Anti-, adalimumab, omalizumab, muromonab-CD3, Rituximab, daclizumab, Herceptin, handkerchief profit pearl Monoclonal antibody, basiliximab and infliximab.

The compound of logical formula (I) as defined herein optionally with one or more following administering drug combinations: ARRY- 162、ARRY-300、ARRY-704、AS-703026、AZD-5363、AZD-8055、BEZ-235、BGT-226、BKM-120、 BYL-719, CAL-101, CC-223, CH-5132799, AP 23573, E-6201, Enzastaurin, GDC-0032, GDC-0068, GDC-0623、GDC-0941、GDC-0973、GDC-0980、GSK-2110183、GSK-2126458、GSK-2141795、MK- 2206, novolimus, OSI-027, piperazine Li Fuxin, PF-04691502, PF-05212384, PX-866, rapamycin, RG- 7167, RO-4987655, RO-5126766, U.S. of department for Buddhist nun, TAK-733, Sibutramine Hydrochloride for Buddhist nun (trametinib), triciribine, UCN-01, WX-554, XL-147, XL-765, azoles Luo Mosi, ZSTK-474.

It is said that in general, compound or the combination of compositions of cytotoxic agent and/or cytostatics with the present invention are used Can play following effect:

(1) produce more preferably reducing tumor growth or even eliminate in tumor side compared with any one medicament individually dosed Effect,

(2) allow to be administered lesser amount of be administered chemotherapeutic agents,

(3) providing chemotherapeutic agent, it is well tolerated by the patient and harmful pharmacology's complication ratio of having is at list One medicament chemotherapy is viewed few with in some other combination treatment,

(4) the various cancers type of the wider array of mammal of therapeutic domain (particularly people) is allowed,

(5) higher response in subject is provided,

(6) time-to-live longer in offer subject compared with the chemotherapeutic treatment of standard,

(7) longer tumour progression time is provided, and/or

(8) compared with the known case that the combination of other cancer agents produces antagonistic effect, obtain at least be used alone Effect that medicament is the best and toleration.

Make cell to radiosensible method

In a different embodiment of the present invention, the compound of the present invention can be used for making cell to radiation-sensitive. That is, before the radiotherapy of cell, compounds for treating cell by the present invention makes the chemical combination of described cell and the unused present invention When thing carries out any treatment, the situation of described cell is compared and is easier to DNA damage and cell death.In an aspect, use The compounds for treating cell of at least one present invention.

Therefore, the present invention also provides for killing the method for cell, wherein by the compound of one or more present invention with conventional X-ray therapy is applied to cell together.

The present invention also provides for making cell be easier to the method that cell death occurs, wherein by one before treating described cell Or cell described in the compounds for treating of the multiple present invention is to cause or inducing cell death.In an aspect, with a kind of or many After planting cell described in the compounds for treating of the present invention, treat by least one compound, at least one method or combinations thereof Described cell is to cause DNA damage thus for suppressing Normocellular function or killing described cell.

In one embodiment, by described cell being killed with at least one DNA damage agent treatment cell.That is, use After the compounds for treating cell of one or more present invention makes described cell by cell death sensitivity, use at least one DNA damage Agent treats described cell to kill described cell.DNA damage agent in the present invention includes but not limited to that chemotherapeutics is (the most suitable Platinum), ionizing radiation (X-ray, ultraviolet radiation), carcinogen and mutagenic agent.

In another embodiment, by with at least one method treatment cell to cause or induced DNA damage is by described Cell is killed.This type of method includes but not limited to: active cell signal transduction pathway (causes DNA when described approach is activated Damage), suppression cellular signal transduction pathways (causing DNA damage when described approach is suppressed) and inducing cell in biology Chemical change (wherein said change causes DNA damage).As limiting examples, capable of inhibiting cell in DNA repair approach, Thus stop repairing and causing the exception of DNA damage in cell to be accumulated of DNA damage.

In one aspect of the invention, carry out radiating or carry out causing in cell before other induction of DNA damage to The compound of the medicine present invention.In another aspect of this invention, lure at other of DNA damage carrying out radiating or carry out to cause cell The compound of the present invention it is administered while leading.In still another aspect of the invention, radiate or carry out to cause the DNA of cell to damage carrying out Other induction of wound is administered the compound of the present invention immediately after starting.

On the other hand, described cell is in vitro.In another embodiment, described cell is in vivo.

As described above, have surprisingly been found that the compound of the present invention effectively suppresses MKNK-1 and therefore can use In treatment or prevention by uncontrolled cell growth, breed and/or survive, unsuitable cellullar immunologic response or unsuitable The disease that cellular inflammation response causes, or with uncontrolled cell growth, breed and/or survive, unsuitable cell Immunne response or the disease of unsuitable cellular inflammation response, especially, the growth of wherein said uncontrolled cell, propagation And/or survival, unsuitable cellullar immunologic response or unsuitable cellular inflammation response are mediated by MKNK-1, such as blood Tumor, solid tumor and/or their transfer, as leukemia and myelodysplastic syndrome, malignant lymphoma, include cerebroma and Brain metastes is at interior head and tumor colli, the breast tumor including non-fire power and small cell lung tumor, stomach Intestinal canal tumour, endocrine tumors, breast tumor and other gynecological tumor, including tumor of kidney, bladder tumor and prostate tumor Urologic neoplasms, cutaneous tumor and sarcoma and/or their transfer.

Therefore, according to it on the other hand, the present invention relates to the compound of the logical formula (I) of as described herein and definition or it stands Body isomer, tautomer, N-oxide, hydrate, solvate or salt particularly pharmaceutically acceptable salt, or Their mixture, it is for treatment or prevention disease as described above.

Therefore, another specific aspect of the present invention be the compound of logical formula (I) as described above or its stereoisomer, Tautomer, N-oxide, hydrate, solvate or salt particularly pharmaceutically acceptable salt, or their mixing Thing is for preventing or treat the purposes of disease.

Therefore, another specific aspect of the present invention be logical formula (I) as described above compound for prepare treatment or The purposes of prophylactic pharmaceutical composition.

Disease mentioned in first two sections be by the growth of uncontrolled cell, breed and/or survive, unsuitable cell The disease that immunne response or unsuitable cellular inflammation response cause, or with uncontrolled cell growth, propagation and/or Survival, unsuitable cellullar immunologic response or the disease of unsuitable cellular inflammation response are especially, wherein said uncontrolled Cell growth, breed and/or survive, unsuitable cellullar immunologic response or unsuitable cellular inflammation response are by MKNK-1 Mediation, such as neoplastic hematologic disorder, solid tumor and/or their transfer, such as leukemia and myelodysplastic syndrome, pernicious pouring Bar tumor, head and tumor colli including cerebroma and brain metastes, include that non-fire power and small cell lung tumor exist In breast tumor, gastroenteric tumor, endocrine tumors, breast tumor and other gynecological tumor, include tumor of kidney, bladder tumor and Prostate tumor is in interior urologic neoplasms, cutaneous tumor and sarcoma and/or their transfer.

In the linguistic context of the present invention, the most as used herein at " unsuitable cellullar immunologic response or inappropriate Cellular inflammation response " linguistic context in, term " unsuitable " be interpreted as preferably representing than normal response more weak or higher also And relevant to the pathology of described disease, cause or cause the response of pathology of described disease.

Preferably, described purposes is the treatment for disease or prevention, and wherein said disease is neoplastic hematologic disorder, solid tumor And/or their transfer.

The method for the treatment of hyperproliferative disorders

The present invention relates to the hyperproliferative disorders of the compound using the present invention and combinations thereof thing treatment mammal Method.Available compound suppress, block, reduce, reduce (etc.) cell proliferation and/or cell division and/or cause wither Die.The method includes being administered to the mammal including people having these needs a certain amount of can effectively treating described disease The compound of the present invention or its pharmaceutically acceptable salt, isomer, polymorph, metabolite, hydrate, solvate or ester Deng.Hyperproliferative disorders includes but not limited to that psoriasis, keloid and other cutaneous hypertrophy, benign prostate increase Raw (BPH), solid tumor such as breast carcinoma, respiratory cancer, the brain cancer, genital cancer, digestive tract cancer, urinary tract cancer, cancer eye, liver Cancer, skin carcinoma, head and neck cancer, thyroid carcinoma, parathyroid carcinoma and the transfer of their far-end.Described disease also include lymphoma, Sarcoma and leukemia.

The example of breast carcinoma includes but not limited to that IDC, ILC, ductal carcinoma in situ and original position are the least Leaf cancer.

The example of respiratory tract cancer include but not limited to small cell lung cancer and nonsmall-cell lung cancer and bronchial adenoma and Pleuropulinonary blastoma.

The example of the brain cancer includes but not limited to brain stem and hypothalamic gliomas, cerebellum and cerebral astrocytoma, marrow mother carefully Born of the same parents' tumor, ependymoma and neuroectodermal tumor and pinealoma.

Genital orgnas,male's tumor includes but not limited to carcinoma of prostate and carcinoma of testis.Tumors of female reproductive organ include but not It is limited to carcinoma of endometrium, cervical cancer, ovarian cancer, cancer of vagina and carcinoma vulvae and sarcoma of uterus.

Digestive tract tumor includes but not limited to anus cancer, colon cancer, colorectal cancer, the esophageal carcinoma, carcinoma of gallbladder, gastric cancer, pancreas Cancer, rectal cancer, carcinoma of small intestine and salivary-gland carcinoma.

Urinary tumor includes but not limited to bladder cancer, carcinoma of penis, renal carcinoma, carcinoma of renal pelvis, carcinoma of ureter, carcinoma of urethra and people Papillary renal carcinoma.

Cancer eye includes but not limited to intraocular melanoma and retinoblastoma.

The example of hepatocarcinoma includes but not limited to hepatocarcinoma (with or without the hepatocarcinoma of fibrolamellar variation), epithelial duct Cancer (intrahepatic cholangiocarcinoma) and Combination hepatocyte cholangiocarcinoma cells.

Skin carcinoma include but not limited to squamous cell carcinoma, Kaposi sarcoma, malignant melanoma, Merkel cell skin cancer with And non-melanoma skin carcinoma.

Head and neck cancer includes but not limited to laryngeal carcinoma, hypopharyngeal cancer, nasopharyngeal carcinoma, oropharynx cancer, lip cancer and oral cancer and squamous epithelial cancer Cell.Lymphoma includes but not limited to that AIDS associated lymphoma, non-Hodgkin lymphoma, cutaneous T cell lymphoma, Hugh Burkitt drench Bar tumor, Hodgkin and central nervous system lymphoma.

Sarcoma includes but not limited to soft tissue sarcoma, osteosarcoma, malignant fibrohistiocytoma, lymphosarcoma and band Myosarcoma.

Leukemia includes but not limited to that acute myeloid leukaemia, acute lymphoblastic leukemia, chronic lymphocytic are white Disorders of blood, chronic myelogenous leukemia and hairy cell leukemia.

These diseases have obtained good sign in the mankind, but are also present in other suckling with similar etiology and move In thing, and can treat by being administered the pharmaceutical composition of the present invention.

Term " treatment (treating) " or the use of " treatment (treatment) " that presents is mentioned in the whole text are conventional , such as manage for the purpose of the situation etc. of the disease or disease of resisting, alleviate, reduce, alleviate, improve such as sarcoma or Nursing individuality.

The method for the treatment of kinases disease

The present invention also provides for the method for treating the disease relevant to abnormal mitogen extracellular kinase activity, described Disease includes but not limited to apoplexy, heart failure, hepatomegaly, cardiomegaly, diabetes, Alzheimer, cystic fibrosis Disease, the symptom of Xenograft rejection, septic shock or asthma.

This type of disease of compounds for treating of the present invention of effective dose can be used, mention including previous Background section that A little diseases (such as cancer).But, this type of cancer of compounds for treating of the available present invention and Other diseases, and and mechanism of action And/or described kinases is unrelated with the relation of described disease.

Phrase " abnormal kinase activity " or " abnormal tyrosine kinase activity " include encoding described kinase whose gene or Any unconventionality expression of the polypeptide of its coding or activity.The example of this type of abnormal activity includes but not limited to described gene or polypeptide Overexpression；Gene amplification；Produce constitutive activity or highly active kinase activity sudden change；Gene mutation, lack, put Change, interpolation etc..

The present invention also provides for the method suppressing kinase activity particularly mitogen extracellular kinase activity, and described method includes The compound of the present invention of effective dosage, including its salt, polymorph, metabolite, hydrate, solvate, prodrug (such as Ester) and its diastereomeric form.Can in cell (the most external) or at mammalian subject in particular for treatment Human patients cell in suppress kinase activity.

The method for the treatment of angiogenesis disease

The present invention also provides for treating the disease relevant to excessive and/or abnormal angiogenesis and the method for disease.

Organism is probably harmful by inappropriate expression and the unconventionality expression of angiogenesis.Many pathological state nothing to do withs (extraneous) growth of blood vessel is correlated with.These include that such as diabetic retinopathy, ischemic retinal vein block And retinopathy of prematurity [Aiello et al., New Engl.J.Med.1994,331,1480；Peer et al., Lab.Invest.1995,72,638], age-related macular degeneration [AMD；See Lopez et al. Invest.Opththalmol.Vis.Sci.1996,37,855], neovascular glaucoma, psoriasis, retrolental increase Restenosis after raw disease, fibrohemangioma, inflammation, rheumatoid arthritis (RA), restenosis, in-stent restenosis, blood vessel transplantation Deng.It addition, the blood supply relevant to cancerous tissue and tumor tissues increases promotes growth, cause quick tumor to increase and turn Move.Additionally, neovascularity and the vasculolymphatic cancerous tumor cell (renegade cells) that is grown to provide the approach of leaving in tumor, promote Enter transfer and cause cancer to spread.Therefore, the compound of the present invention can be used to treat and/or prevent any mentioned above Angiogenesis disease, such as by suppression and/or minimizing vascularization；Suppress, block, reduce, reduce (etc.) endotheliocyte Propagation or other type relevant to angiogenesis, and cause cell death or the apoptosis of this type of cell.

Dosage and administration

The standard of the compound for treating hyperproliferative disorders and angiogenesis disease is evaluated in fact based on known being used for Test room technology, by standard toxicity test and by the standard for determining the treatment to disease mentioned above in mammal Pharmacology test, and by these results and the result being used for treating the known drug of these diseases are compared, can hold Change places and determine the effective dose of the compound for treating each present invention expecting indication.Controlling in one of these diseases Given in treatment, the amount of the active component of medicine can be according to considering as follows and great changes will take place: the particular compound used and dosage Unit, administering mode, the course for the treatment of, the age of subject and sex and the nature and extent of condition being treated.

The total amount of active component to be administered is typically about 0.001mg/kg-about 200mg/kg body weight/day, and preferably from about 0.01mg/kg-about 20mg/kg body weight/day.The most useful dosage regimen can be once a day to three times be administered to every four The administration of Zhou Yici.It addition, " withdrawal time " (within certain a period of time, wherein not giving Patient drug) for pharmacological effect and Whole machine balancing between toleration is probably favourable.Unit dose can include about 0.5mg-about 1500mg active component, and Can be administered one or more times daily, or less than being administered once a day.By including intravenous, intramuscular, subcutaneous and the intestines and stomach The ADD being injected at outward interior injection and use infusion techniques administration can be preferably that 0.01-200mg/kg is overall Weight.Averagely every day, rectal dosage regimen was preferably 0.01-200mg/kg TBW.Averagely every day, vaginal dosage scheme was preferably 0.01-200mg/kg TBW.Averagely topical dosage regimen every day is preferably the most once a day to four administration 0.1-200mg.Thoroughly Skin concentration is preferably the concentration required for maintaining the daily dosage of 0.01-200mg/kg.Averagely every day, inhalation dose scheme was preferred For 0.01-100mg/kg TBW.

The concrete initial dose of certain each patient and maintenance dose scheme can change according to following factor: clinical The character of disease determined by diagnostician and severity, the particular compound used patient active, described age and Holistic health, administration time, route of administration, the discharge rate of medicine, drug regimen etc..Therefore, the compound of the present invention Or the desired therapeutic modality of its pharmaceutically acceptable salt or ester or compositions and be administered quantity can be by those skilled in the art Conventional therapeutic test is utilized to determine.

Preferably, the disease that described method is targeted is neoplastic hematologic disorder, solid tumor and/or their transfer.

The compound of the present invention is used especially for treatment and prevents (i.e. prevention) growth and metastasis of tumours, particularly accept or Do not accept all indications and the growth and metastasis of tumours of the solid tumor in stage of the pre-treatment of described tumor growth.

Concrete pharmacological property or the method for testing of pharmaceutical properties are to well known to a person skilled in the art.

Embodiment described herein that test experiments is for illustrating the present invention and the invention is not restricted to provided reality Execute example.

Bioassay:

Embodiment is tested one or more times by selected bioassay.When testing more than once, data report is flat Average or median, wherein:

● meansigma methods, also known as arithmetic mean of instantaneous value, represent the sum of value obtained divided by the number of times tested, and

● median represents the median of numerical value group when with ascending order or descending.If the numerical value in data set Number is odd number, then median is middle numerical value.If the number of the numerical value in data set is even number, then median is two The arithmetic mean of instantaneous value of the numerical value of individual centre.

Synthetic example is one or more times.When synthesizing more than once, the data from bioassay represent that utilization is by surveying The data set that tries one or more synthesis batch and obtain and the meansigma methods that calculates or median.

MKNK1 kinase assays

The MKNK1TR-FRET as described in paragraphs below is utilized to measure the MKNK1-suppression of the compound quantifying the present invention Activity.

From Carna Biosciences (production number 02-145) buy glutathione-S-transferase (GST, N-end) and The recombination fusion protein of people overall length MKNK1 (amino acid/11-424 and the T344D of preserving number BAA 19885.1) is also used as enzyme, institute State recombination fusion protein express in the insect cell using baculovirus expression system and pass through glutathione agarose affinity Chromatograph carrys out purification.Use biotinylated peptide biotin-Ahx-IKKRKLTRRKSLKG (the C-end of amide form thereof) as kinases The substrate of reaction, it is purchased from such as Biosyntan company (Berlin-Buch, Germany).

For measuring, the test-compound of 50nL 100 times of concentrated solution pipettors in DMSO are sucked the low appearance of black Measuring 384 hole titer plate (Greiner Bio-One, Frickenhausen, Germany), the MKNK1 adding 2 μ L surveys in aqueous Determine buffer [50mM HEPES pH 7.5,5mM magnesium chloride, 1.0mM dithiothreitol, DTT, 0.005% (v/v) Nonidet-P40 (Sigma) solution in], and mixture is hatched 15min so that test-compound is before starting kinase reaction at 22 DEG C It is incorporated into this enzyme in advance.Then, by add 3 μ L adenosine triphosphate (ATP, 16.7 μMs=> in 5 μ L measure in volumes final Concentration is 10 μMs) and substrate (0.1 μM=> ultimate density in 5 μ L test volume be 0.06 μM) in measuring in buffer Solution starts kinase reaction, and gained mixture is hatched the response time of 45min at 22 DEG C.Activity according to enzyme batch is come The concentration of regulation MKNK1, and properly select so that measuring and being in the range of linearity, typical concentration is the scope of 0.05 μ g/ml. By add 5 μ L TR-FRET detectable (5nM streptavidin-XL665 [Cisbio Bioassays, Codolet, France] and from 1nM anti-ribosomal protein S6 (the pSer236)-antibody [#44921G] of Invitrogen and 1nM The ProteinG [Perkin-Elmer, production number AD0071] of LANCE EU-W1024 labelling) in EDTA aqueous solution (100mM The bovine serum albumin pH 7.5 in 50mM HEPES of EDTA, 0.1% (w/v)) in solution terminate reaction.

Gained mixture is hatched 1h so that being formed multiple between biotinylation peptide and the detectable of phosphorylation at 22 DEG C Compound.The amount of phosphorylated substrate is evaluated subsequently by measuring the resonance energy transfer of-XL from Eu-chelate to streptavidin. Therefore, TR-FRET reader, such as (BMG Labtechnologies, Offenburg, Germany) or Viewlux are utilized (Perkin-Elmer) measure after 350nm excites, in the fluorescent emission of 620nm and 665nm.665nm and 622nm sends out The ratio penetrated is used as measuring of the amount of phosphorylated substrate.By data normalization, (enzyme reaction=0% suppression of no inhibitor, has institute Other is had to measure component and suppress without enzyme=100%).Generally, test-compound on identical microtitration plate with 11 kinds not Test with concentration, each concentration determination two value, and use and calculate IC by 4 parameter fittings₅₀Value, described concentration is 20 μM to 0.1nM (20 μMs, 5.9 μMs, 1.7 μMs, 0.51 μM, 0.15 μM, 44nM, 13nM, 3.8nM, 1.1nM,

0.33nM and 0.1nM, before the assay, is divided by 1:3.4 serial dilution in the level of the dense DMSO solution of 100 times Do not prepare this dilution series).

Table 1:MKNK1IC₅₀

Embodiment	MKNK1IC₅₀[nM]
		1	4
3	4
		4	4
6	7
		8	6
9	7
		10	8

11	12
		12	11
13	9
		15	12
16	16
		17	8
18	24
		19	16
21	15
		22	25
23	25
		25	39
27	36
		28	59
29	98
		30	227
31	22
		32	28
33	10
		34	10

MKNK1 kinases height ATP measures

The compound utilizing the mensuration of the MKNK1 height ATP based on TR-FRET as described in paragraphs below to quantify the present invention exists Its with MKNK1 preincubate after MKNK1-inhibitory activity under high ATP.

For measuring, the test-compound of 50nL 100 times of concentrated solution pipettors in DMSO are sucked the low appearance of black Measuring 384 hole titer plate (Greiner Bio-One, Frickenhausen, Germany), the MKNK1 adding 2 μ L surveys in aqueous Determine buffer [50mM HEPES pH 7.5,5mM magnesium chloride, 1.0mM dithiothreitol, DTT, 0.005% (v/v) Nonidet-P40 (Sigma) solution in], and mixture is hatched 15min so that test-compound is before starting kinase reaction at 22 DEG C It is incorporated into this enzyme in advance.Then, by add 3 μ L adenosine triphosphate (ATP, 3.3mM=> in 5 μ L measure in volumes the denseest Degree is for 2mM) and the substrate (0.1 μM=> ultimate density in 5 μ L test volume be 0.06 μM) solution in mensuration buffer Start kinase reaction, and gained mixture is hatched at 22 DEG C the response time of 30min.Activity according to enzyme batch regulates The concentration of MKNK1, and properly select so that measuring and being in the range of linearity, typical concentration is the scope of 0.003 μ g/mL.Logical Cross add 5 μ L TR-FRET detectable (5nM streptavidin-XL665 [Cisbio Bioassays, Codolet, France] and from 1nM anti-ribosomal protein S6 (the pSer236)-antibody [#44921G] of Invitrogen and 1nM The ProteinG [Perkin-Elmer, production number AD0071] of LANCE EU-W1024 labelling) in EDTA aqueous solution (100mM The bovine serum albumin pH 7.5 in 50mM HEPES of EDTA, 0.1% (w/v)) in solution terminate reaction.

Gained mixture is hatched 1h so that being formed multiple between biotinylation peptide and the detectable of phosphorylation at 22 DEG C Compound.The amount of phosphorylated substrate is evaluated subsequently by measuring the resonance energy transfer of-XL from Eu-chelate to streptavidin. Therefore, utilize TR-FRET reader, such as Rubystar (BMG Labtechnologies, Offenburg, Germany) or Viewlux (Perkin-Elmer) measures after 350nm excites, in the fluorescent emission of 620nm and 665nm.665nm and The ratio of the transmitting of 622nm is used as measuring of the amount of phosphorylated substrate.By data normalization, (enzyme reaction=0% of no inhibitor presses down System, has all other and measures component and suppress without enzyme=100%).Generally, test-compound is on identical microtitration plate Test with 11 kinds of variable concentrations, each concentration determination two value, and use and calculate IC by 4 parameter fittings₅₀Value, described Concentration be 20 μMs to 0.1nM (such as, 20 μMs, 5.9 μMs, 1.7 μMs, 0.51 μM, 0.15 μM, 44nM, 13nM, 3.8nM, 1.1nM, 0.33nM and 0.1nM, before the assay, prepares this dilution respectively by serial dilution in the level at the dense DMSO solution of 100 times Series, concentration can change according to pipettor used accurately).

Table 2:MKNK1 height ATP IC₅₀

Embodiment	MKNK1 height ATP IC₅₀[nM]
		1	6
2	11
		3	12
4	13
		5	18
6	18
		7	19
8	20
		9	20
10	23
		11	24
12	25
		13	25
14	27
		15	30
16	32
		17	36
18	37
		19	38
20	41
		21	44
22	53
		23	53
24	55
		25	91
26	93
		27	75
28	114
		29	124

30	239
		31	49
32	53
		33	15
34	11

CDK2/CycE kinase assays

The CDK2/CycE TR-FRET as described in paragraphs below is utilized to measure the CDK2/ of the compound quantifying the present invention CycE-inhibitory activity.

From ProQinase GmbH (Freiburg, Germany) buy GST and the recombination fusion protein of people CDK2 and GST and the recombination fusion protein of people CycE, described recombination fusion protein is all expressed and by gluathione in insect cell (Sf9) Peptide-agarose affinity chromatography carrys out purification.Use biotinylated peptide biotin-Ttds-YISPLKSPYKISEG (amide form thereof C-end) as the substrate of kinase reaction, it is purchased from such as JERINI peptide scientific & technical corporation (Berlin, Germany).

For measuring, the test-compound of 50nL 100 times of concentrated solution pipettors in DMSO are sucked the low appearance of black Measure 384 hole titer plate (Greiner Bio-One, Frickenhausen, Germany), add the CDK2/CycE of 2 μ L in water Property measure buffer [50mM Tris/ hydrochloric acid pH8.0,10mM magnesium chloride, 1.0mM dithiothreitol, DTT, 0.1mM sodium vanadate, 0.01% (v/v) Nonidet-P40 (Sigma)] in solution, and mixture is hatched 15min so that testedization at 22 DEG C Compound was incorporated into this enzyme before starting kinase reaction in advance.Then, by add 3 μ L adenosine triphosphate (ATP, 16.7 μMs=> The ultimate density measured in volumes in 5 μ L is 10 μMs) and substrate (1.25 μMs=> ultimate density in 5 μ L test volume is 0.75 μM) start kinase reaction in the solution measured in buffer, and gained mixture is hatched at 22 DEG C the reaction of 25min Time.Activity according to enzyme batch regulates the concentration of CDK2/CycE, and properly selects so that measuring and being in the range of linearity, Typical concentration is the scope of 130ng/mL.By adding TR-FRET detectable (0.2 μM of streptavidin-XL665 of 5 μ L [Cisbio Bioassays, Codolet, France] and the 1nM anti-RB (pSer807/ from BD Pharmingen PSer811)-antibody [#558389] and the anti-mouse IgG antibody [Perkin-of 1.2nM LANCE EU-W1024 labelling Elmer, production number AD0077, as an alternative, can use the terbium-cryptate-labelling from Cisbio Bioassays Anti-mouse IgG antibody]) in EDTA aqueous solution (100mM EDTA, 0.2% (w/v) in 100mM HEPES/ sodium hydroxide Bovine serum albumin pH 7.0) in solution terminate reaction.

Gained mixture is hatched 1h so that being formed multiple between biotinylation peptide and the detectable of phosphorylation at 22 DEG C Compound.The amount of phosphorylated substrate is evaluated subsequently by measuring the resonance energy transfer of-XL from Eu-chelate to streptavidin. Therefore, utilize TR-FRET reader, such as Rubystar (BMG Labtechnologies, Offenburg, Germany) or Viewlux (Perkin-Elmer) measures after 350nm excites, in the fluorescent emission of 620nm and 665nm.665nm and The ratio of the transmitting of 622nm is used as measuring of the amount of phosphorylated substrate.By data normalization, (enzyme reaction=0% of no inhibitor presses down System, has all other and measures component and suppress without enzyme=100%).Generally, test-compound is on identical microtitration plate Test with 11 kinds of variable concentrations, each concentration determination two value, and use and calculate IC by 4 parameter fittings₅₀Value, described Concentration be 20 μMs to 0.1nM (20 μMs, 5.9 μMs, 1.7 μMs, 0.51 μM, 0.15 μM, 44nM, 13nM, 3.8nM, 1.1nM, 0.33nM And 0.1nM, before the assay, the level of the dense DMSO solution of 100 times prepares this dilution system respectively by 1:3.4 serial dilution Row).

PDGFR beta kinase measures

The PDGFR β suppression utilizing the PDGFR β HTRF as described in paragraphs below to measure the compound quantifying the present invention is lived Property.

As kinases, use from Proqinase [Freiburg i.Brsg., Germany] buy containing people PDGFR β's The GST-His fusion protein of C-end fragment (aminoacid 561-1106), it is expressed and by affinity color in insect cell [SF9] Spectrum carrys out purification.Use from Cis Biointernational (Marcoule, France) the poly-Glu of biotinylation, Tyr (4: 1) copolymer (#61GT0BLA) is as the substrate of kinase reaction.

For measuring, the test-compound of 50nL 100 times of concentrated solution pipettors in DMSO are sucked the low appearance of black Measure 384 hole titer plate (Greiner Bio-One, Frickenhausen, Germany), add the PDGFR β of 2 μ L in aqueous Measure buffer [50mM HEPES/ sodium hydroxide pH 7.5,10mM magnesium chloride, 2.5mM dithiothreitol, DTT, 0.01% (v/v) Triton-X100 (Sigma)] in solution, and mixture is hatched 15min so that test-compound swashs starting at 22 DEG C This enzyme it is incorporated in advance before enzyme reaction.Then, by add 3 μ L adenosine triphosphate (ATP, 16.7 μMs=> in 5 μ L measure bodies Ultimate density in Ji is 10 μMs) and substrate (2.27 μ g/mL=> ultimate density in 5 μ L test volume are 1.36 μ g/mL [about 30nM]) start kinase reaction in the solution measured in buffer, and gained mixture is hatched the anti-of 25min at 22 DEG C Between Ying Shi.Regulate the PDGFR β concentration in mensuration according to enzyme batch activity, and suitably select so that measuring in the range of linearity In, typical enzyme concentration is about the scope (ultimate density in 5 μ L measure volume) of 125pg/ μ L.By adding 5 μ L's HTRF detectable (200nM streptavidin-XLent [Cis Biointernational] and 1.4nM PT66-Eu-chelating (the anti-phosphotyrosine antibody from the europium-chelate labels of Perkin Elmer [can also use from Cis thing The PT66-Tb-cryptate of Biointernational substitutes PT66-Eu-chelate]) in EDTA aqueous solution (100mM EDTA, 0.2% (w/v) the bovine serum albumin pH 7.5 in 50mM HEPES/ sodium hydroxide) in solution terminate instead Should.

By the mixture of gained 22 DEG C hatch 1h so that biotinylated Phosphorylated Peptide and streptavidin-XLent and PT66-Eu-chelate combines.Subsequently by measuring the resonance energy transfer of-XLent from PT66-Eu-chelate to streptavidin Evaluate the amount of phosphorylated substrate.Therefore, utilize HTRF reader, such as Rubystar (BMG Labtechnologies, Offenburg, Germany) or Viewlux (Perkin-Elmer) measure after 350nm excites, at 620nm and 665nm Fluorescent emission.The ratio of the transmitting of 665nm and 622nm is used as measuring of the amount of phosphorylated substrate.By data normalization (unrestraint Enzyme reaction=0% suppression of agent, has all other and measures component and suppress without enzyme=100%).Generally, test-compound Identical titer plate is tested with 10 kinds of variable concentrations, each concentration determination two value, and use by 4 parameter fittings Calculate IC₅₀Value, described concentration be 20 μMs to 0.1nM (20 μMs, 6.7 μMs, 2.2 μMs, 0.74 μM, 0.25 μM, 82nM, 27nM, 9.2nM, 3.1nM and 1nM, before the assay, prepared by 1:3 serial dilution in the level of the dense stock solution of 100 times respectively This dilution series).

Fyn kinase assays

With the people of His6-label kinases territory of recombinating, the C-end of people T-Fyn is used as kinases, and it is at baculovirus infection Insect cell (purchased from Invitrogen, P3042) is expressed.Use biotinylated peptide biotin-KVEKIGEGTYGVV (acyl The C-end of amine form) as the substrate of kinase reaction, its be purchased from such as Biosynthan GmbH company (Berlin-Buch, Germany)。

For measuring, the test-compound of 50nL 100 times of concentrated solution pipettors in DMSO are sucked the low appearance of black Measuring 384 hole titer plate (Greiner Bio-One, Frickenhausen, Germany), the T-Fyn adding 2 μ L surveys in aqueous Determine buffer [25mM Tris/ hydrochloric acid pH 7.2,25mM magnesium chloride, 2mM dithiothreitol, DTT, 0.1% (w/v) bovine serum albumin In vain, 0.03% (v/v) Nonidet-P40 (Sigma)] in solution, and mixture is hatched 15min so that tested at 22 DEG C Compound was incorporated into this enzyme before starting kinase reaction in advance.Then, by add 3 μ L adenosine triphosphate (ATP, 16.7 μMs => ultimate density that measures in volumes in 5 μ L is 10 μMs) and substrate (2 μMs=> ultimate density in 5 μ L test volume is 1.2 μMs) start kinase reaction in the solution measured in buffer, and gained mixture is hatched at 22 DEG C the reaction of 60min Time.Activity according to enzyme batch regulates the concentration of Fyn, and properly selects so that measuring and being in the range of linearity, typically Concentration is 0.13nM.By add 5 μ L HTRF detectable (0.2 μM of streptavidin-XL [Cisbio Bioassays, Codolet, France] and 0.66nM PT66-Eu-chelate (the anti-phosphorus from the europium-chelate labels of Perkin Elmer Acid phosphotyrosine antibody [can also use the PT66-Tb-cryptate from Cisbio Bioassays to substitute PT66-Eu- Chelate]) in EDTA aqueous solution (125mM EDTA, 0.2% (w/v) bovine serum albumin in 50mM HEPES/ sodium hydroxide White pH 7.0) in solution terminate reaction.

The mixture of gained is hatched 1h so that biotinylation Phosphorylated Peptide and streptavidin-XL and PT66-at 22 DEG C Eu-chelate combines.Phosphorus is evaluated subsequently by the resonance energy transfer of measurement-XL from PT66-Eu-chelate to streptavidin The amount of acidifying substrate.Therefore, utilize HTRF reader, such as Rubystar (BMG Labtechnologies, Offenburg, Germany) or Viewlux (Perkin-Elmer) measures after 350nm excites, the fluorescence at 620nm and 665nm is sent out Penetrate.The ratio of the transmitting of 665nm and 622nm is used as measuring of the amount of phosphorylated substrate.By data normalization, (enzyme of no inhibitor is anti- Answer=0% suppression, there is all other and measure component and suppress without enzyme=100%).Generally, test-compound is identical micro- Test with 10 kinds of variable concentrations on titer plate, each concentration determination two value, and use and calculated by 4 parameter fittings IC₅₀Value, described concentration be 20 μMs to 0.1nM (20 μMs, 6.7 μMs, 2.2 μMs, 0.74 μM, 0.25 μM, 82nM, 27nM, 9.2nM, 3.1nM and 1nM, before the assay, prepares this dilution by 1:3 serial dilution in the level of the dense stock solution of 100 times respectively Series).

Flt4 kinase assays

The Flt4 suppression utilizing the Flt4TR-FRET as described in paragraphs below to measure the compound quantifying the present invention is lived Property.

As kinases, use the C-containing people Flt4 bought from Proqinase [Freiburg i.Brsg., Germany] The GST-His fusion protein of end fragment (aminoacid 799 1298), it is expressed in insect cell [SF9] and passes through affinity chromatography Carry out purification.(the C-end of amide form thereof, is purchased to use biotinylated peptide biotin-Ahx-GGEEEEYFELVKKKK Biosyntan, Berlin-Buch, Germany) as the substrate of kinase reaction.

For measuring, the test-compound of 50nL 100 times of concentrated solution pipettors in DMSO are sucked the low appearance of black Measuring 384 hole titer plate (Greiner Bio-One, Frickenhausen, Germany), the Flt4 adding 2 μ L surveys in aqueous Determine buffer [25mM HEPES pH 7.5,10mM magnesium chloride, 2mM dithiothreitol, DTT, 0.01% (v/v) Triton-X100 (Sigma), 0.5mM EGTA and 5mM β-phosphoglycerol] in solution, and mixture is hatched 15min at 22 DEG C so that Test-compound was incorporated into this enzyme before starting kinase reaction in advance.Then, by add 3 μ L adenosine triphosphate (ATP, 16.7 μMs=> ultimate density that measures in volumes in 5 μ L is 10 μMs) and substrate (1.67 μMs=> in 5 μ L test volume Final concentration of 1 μM) start kinase reaction in the solution measured in buffer, and gained mixture is hatched 45min at 22 DEG C Response time.Activity according to enzyme batch regulates the Flt4 concentration in mensuration, and suitably selects so that measuring linearly In scope, typical enzyme concentration is about the scope (ultimate density in 5 μ L measure volume) of 120pg/ μ L.By adding 5 μ L HTRF detectable (200nM streptavidin-XL665 [Cis Biointernational] and 1nM PT66-Tb-is cave-shaped Compound (the anti-phosphotyrosine from the terbium-cryptate labelling of Cisbio Bioassays (Codolet, France) Antibody)) in EDTA aqueous solution (50mM EDTA, 0.2% (w/v) the bovine serum albumin pH 7.5 in 50mM HEPES) Solution terminate reaction.

By the mixture of gained 22 DEG C hatch 1h so that biotinylated Phosphorylated Peptide and streptavidin-XL665 and PT66-Tb-cryptate combines.Subsequently by measuring being total to of-XL665 from PT66-Tb-cryptate to streptavidin Shake and can shift the amount evaluating phosphorylated substrate.Therefore, HTRF reader, such as Rubystar (BMG are utilized Labtechnologies, Offenburg, Germany) or Viewlux (Perkin-Elmer) measure and excite it at 350nm After, in the fluorescent emission of 620nm and 665nm.The ratio of the transmitting of 665nm and 622nm is used as measuring of the amount of phosphorylated substrate.Will (enzyme reaction=0% suppression of no inhibitor has all other and measures component and press down without enzyme=100% data normalization System).Generally, test-compound is tested with 10 kinds of variable concentrations in identical titer plate, each concentration determination two value, and And use calculates IC by 4 parameter fittings₅₀Value, described concentration be 20 μMs to 0.1nM (20 μMs, 6.7 μMs, 2.2 μMs, 0.74 μM, 0.25 μM, 82nM, 27nM, 9.2nM, 3.1nM and 1nM, before the assay, pass through 1:3 in the level of the dense stock solution of 100 times Serial dilution prepares this dilution series respectively).

TrkA kinase assays

The TrkA HTRF as described in paragraphs below is utilized to measure the TrkA inhibitory activity of the compound quantifying the present invention.

As kinases, use the C-containing people TrkA bought from Proqinase [Freiburg i.Brsg., Germany] The GST-His fusion protein of end fragment (aminoacid 443 796), it is expressed in insect cell [SF9] and passes through affinity chromatography Carry out purification.Use the poly-Glu of biotinylation, Tyr (4:1) from Cis Biointernational (Marcoule, France) Copolymer (#61GT0BLA) is as the substrate of kinase reaction.

For measuring, the test-compound of 50nL 100 times of concentrated solution pipettors in DMSO are sucked the low appearance of black Measuring 384 hole titer plate (Greiner Bio-One, Frickenhausen, Germany), the TrkA adding 2 μ L surveys in aqueous Determine buffer [8mM MOPS/ hydrochloric acid pH 7.0,10mM magnesium chloride, 1mM dithiothreitol, DTT, 0.01% (v/v) NP-40 (Sigma), 0.2mMEDTA] in solution, and mixture is hatched 15min so that test-compound swashs starting at 22 DEG C This enzyme it is incorporated in advance before enzyme reaction.Then, by add 3 μ L adenosine triphosphate (ATP, 16.7 μMs=> in 5 μ L measure bodies Ultimate density in Ji is 10 μMs) and substrate (2.27 μ g/mL=> ultimate density in 5 μ L test volume are 1.36 μ g/ml [about 30nM]) start kinase reaction in the solution measured in buffer, and gained mixture is hatched the anti-of 60min at 22 DEG C Between Ying Shi.Activity according to enzyme batch regulates the TrkA concentration in mensuration, and suitably selects so that measuring in the range of linearity In, typical enzyme concentration is about the scope (ultimate density in 5 μ L measure volume) of 20pg/ μ L.By adding the HTRF of 5 μ L (30nM streptavidin-XL665 [Cis Biointernational] and 1.4nM PT66-Eu-chelate (come detectable [can also use from Cis from the anti-phosphotyrosine antibody of the europium-chelate labels of Perkin Elmer The PT66-Tb-cryptate of Biointernational substitutes PT66-Eu-chelate]) in EDTA aqueous solution (100mM EDTA, 0.2% (w/v) the bovine serum albumin pH 7.5 in 50mM HEPES/ sodium hydroxide) in solution terminate instead Should.

By the mixture of gained 22 DEG C hatch 1h so that biotinylated Phosphorylated Peptide and streptavidin-XL665 and PT66-Eu-chelate combines.Subsequently by measuring the resonance energy transfer of-XL665 from PT66-Eu-chelate to streptavidin Evaluate the amount of phosphorylated substrate.Therefore, utilize HTRF reader, such as Rubystar (BMG Labtechnologies, Offenburg, Germany) or Viewlux (Perkin-Elmer) measure after 350nm excites, at 620nm and 665nm Fluorescent emission.The ratio of the transmitting of 665nm and 622nm is used as measuring of the amount of phosphorylated substrate.By data normalization (unrestraint Enzyme reaction=0% suppression of agent, has all other and measures component and suppress without enzyme=100%).Generally, test-compound Identical titer plate is tested with 10 kinds of variable concentrations, each concentration determination two value, and use by 4 parameter fittings Calculate IC₅₀Value, described concentration be 20 μMs to 0.1nM (20 μMs, 6.7 μMs, 2.2 μMs, 0.74 μM, 0.25 μM, 82nM, 27nM, 9.2nM, 3.1nM and 1nM, before the assay, prepared by 1:3 serial dilution in the level of the dense stock solution of 100 times respectively This dilution series).

AlphaScreen SureFire eIF4E Ser209 phosphorylation assay

AlphaScreen SureFire eIF4E Ser209 phosphorylation assay is used for measuring endogenous eIF4E in cell Phosphorylation in lysate.AlphaScreen SureFire technology can measure the phosphorylation egg in cell lysates In vain.In this mensuration, AlphaScreen donor and Acceptor beads capture and only exist in analyte (p-eIF4E Ser209) The sandwich antibody complex of lower formation, makes them in close proximity.Donor microballon excite the release causing singlet oxygen atom, its Trigger the energy transfer cascade in Acceptor beads, produce the light emission of 520-620nm.

The Surefire EIF4e Alphascreen stimulated with 20%FCS in A549 cell

For measuring, use the AlphaScreen SureFire p-eIF4E Ser209 being all from Perkin Elmer 10K assay kit and AlphaScreen ProteinA test kit (for 10K measuring point).

At first day, in 96 orifice plates, it is inoculated in growth medium with every hole 100 μ L (there is stable valley glutamine DMEM/Hams ' F12,10%FCS) in 50.000A549 cell, and hatch at 37 DEG C.After cell attachment, by culture medium Become starvation media (DMEM, 0.1%FCS, do not have glucose, has glutamine, is supplemented with 5g/L maltose).Second My god, test-compound serial dilution in 50 μ L starvation media, and finally DMSO concentration is 1%, and add it to A549 cell in test board, ultimate density scope is that 10 μMs of up to 10nM are low according to the concentration of test-compound.At Jiang The cell of reason hatches 2h at 37 DEG C.37 μ L FCS are added to hole (=final FCS concentration 20%), continue 20min.Then remove Culture medium, and dissolve cell by adding 50 μ L Cell lysis buffer.Then, swing plate 10min on plate shaker. After 10min cell dissolution time, the lysate of 4 μ L is transferred to 384 orifice plates, and (Proxiplate, from Perkin And add 5 μ L and add activation buffer solution mixture containing the reaction buffer of AlphaScreen Acceptor beads Elmer),.With TopSeal-A glued membrane sealing plate, at room temperature, softly shakes 2h on plate shaker.Hereafter, under sheen, add 2 μ L to have The dilution buffer of AlphaScreen donor microballon, and again use TopSeal-A glued membrane sealing plate, and cover with paper tinsel.Carry out Additionally 2h is hatched and is the most softly shaken.Then, there is the EnVision reader of AlphaScreen program (Perkin Elmer) measures plate.Measure each data point (diluted chemical compound) in triplicate.

IC is measured by 4-parameter fitting₅₀Value.

It is aobvious for those skilled in the art that applicable reagent can be used to carry out the kinase whose mensuration of other MKNK-1 similarly And be clear to.

Therefore, the compound of the present invention effectively suppress one or more MKNK-1 kinases and be consequently adapted to treatment or prevention by The growth of uncontrolled cell, breed and/or survive, unsuitable cellullar immunologic response or unsuitable cellular inflammation response are drawn Rise disease, especially, wherein said uncontrolled cell growth, breed and/or survive, unsuitable cellullar immunologic response Or unsuitable cellular inflammation response is mediated by MKNK-1, more particularly, wherein said by the growth of uncontrolled cell, The disease that propagation and/or survival, unsuitable cellullar immunologic response or unsuitable cellular inflammation response cause be neoplastic hematologic disorder, Solid tumor and/or their transfer, such as leukemia and myelodysplastic syndrome, malignant lymphoma, include cerebroma and brain Transfer is at interior head and tumor colli, the breast tumor including non-fire power and small cell lung tumor, gastrointestinal Road tumor, endocrine tumors, breast tumor and other gynecological tumor, secreting including tumor of kidney, bladder tumor and prostate tumor Urinary system tumor, cutaneous tumor and sarcoma and/or their transfer.

Claims

1. the compound of logical formula (I) or its stereoisomer, tautomer or pharmaceutically acceptable salt, or theirs is mixed Compound:

Wherein:

Represent:

Group；

R1 represents linear-C₁-C₆-alkyl-, branched-C₃-C₆-alkyl-or-C₃-C₆-cycloalkyl-, it is optionally selected from Following substituent group replacement once or independently of each other replaces repeatedly:

Halogen atom ,-CN, C₁-C₆-alkyl-, aryl-,-OH, C₁-C₆-alkoxyl-；

R2 represents hydrogen atom；

R3 represents selected from following substituent group:

Halogen atom ,-CN, C₁-C₆-alkyl-, C₁-C₆-haloalkyl-,-OH, C₁-C₆-alkoxyl-；

R4 represents selected from following substituent group:

Hydrogen atom；

R5 represents:

Or:

-selected from following substituent group: C₁-C₆-alkyl-, C₁-C₆-haloalkyl-, C₃-C₁₀-cycloalkyl-, C₃-C₁₀-cycloalkyl- C₁-C₆-alkyl-；

Or:

The carbon atom of-the nitrogen-atoms that is connected with it and R1 be collectively forming 3 yuan to 7 ring secondary amine,

N represents the integer of 0,1,2,3,4 or 5.

2. the compound of claim 1 or its stereoisomer, tautomer or pharmaceutically acceptable salt, or they Mixture, wherein:

Represent:

Group；

C₁-C₆-alkyl-, aryl-,-OH, C₁-C₆-alkoxyl-；

R2 represents hydrogen atom；

R3 represents selected from following substituent group:

R4 represents selected from following substituent group:

Hydrogen atom；

R5 represents:

Or:

N represents the integer of 0,1,2,3,4 or 5.

3. the compound of claim 1 or its stereoisomer, tautomer or pharmaceutically acceptable salt, or they Mixture is wherein:

Represent:

Group；

R1 represents linear-C₁-C₆-alkyl-or branched-C₃-C₆-alkyl-, it is optionally selected from following substituent group and replaces Once or independently of each other replace repeatedly:

C₁-C₆-alkyl-, aryl-,-OH, C₁-C₆-alkoxyl-；

R2 represents hydrogen atom；

R3 represents selected from following substituent group:

R4 represents selected from following substituent group:

Hydrogen atom；

R5 represents:

Or:

The carbon atom of-the nitrogen-atoms that is connected with it and R1 is collectively forming 3 yuan to 7 ring secondary amine；

N represents the integer of 0 or 1.

4. the compound of claim 1 or its stereoisomer, tautomer or pharmaceutically acceptable salt, or they Mixture, wherein:

Represent:

Group；

R1 represents linear-C₁-C₅-alkyl-or branched-C₃-C₅-alkyl-, it is optionally selected from following substituent group and replaces Once or independently of each other replace repeatedly:

C₁-C₆-alkyl-or aryl-；

R2 represents hydrogen atom；

R3 represents selected from following substituent group:

R4 represents selected from following substituent group:

Hydrogen atom；

R5 represents:

Or:

N represents the integer of 0 or 1.

5. the compound of claim 1 or its stereoisomer, tautomer or pharmaceutically acceptable salt, or they Mixture, wherein:

Represent:

Group；

R1 represents linear-C₁-C₅-alkyl-, it is optionally substituted base and replaces once, and described substituent group is:

Aryl-；

R2 represents hydrogen atom；

R3 represents selected from following substituent group:

Halogen atom, C₁-C₆-alkoxyl-；

R4 represents hydrogen atom；

R5 represents:

Or:

N represents the integer of 0 or 1.

6. the compound any one of claim 1-5 or its stereoisomer, tautomer or pharmaceutically acceptable salt, Or their mixture, it is selected from:

3-(1-benzofuran-2-base)-6-[2-(morpholine-2-yl) ethyoxyl] imidazo [1,2-b] pyridazine；

3-(4-methoxyl group-1-benzofuran-2-base)-6-[(2R)-morpholine-2-ylmethoxy] imidazo [1,2-b] pyridazine；

3-(1-benzofuran-2-base)-6-(morpholine-2-ylmethoxy) imidazo [1,2-b] pyridazine；

N-(3-{ [3-(1-benzofuran-2-base) imidazo [1,2-b] pyridazine-6-base] epoxide } propyl group)-2,2-dimethyl propylene Alkane-1-amine；

3-(5-methoxyl group-1-benzofuran-2-base)-6-[(2R)-morpholine-2-ylmethoxy] imidazo [1,2-b] pyridazine；

2-{ [3-(1-benzofuran-2-base) imidazo [1,2-b] pyridazine-6-base] epoxide }-N-(Cvclopropvlmethvl) ethamine；

3-(1-benzofuran-2-base)-6-[(2R)-morpholine-2-ylmethoxy] imidazo [1,2-b] pyridazine；

3-(1-benzofuran-2-base)-6-{2-[(3R)-morpholine-3-base] ethyoxyl } imidazo [1,2-b] pyridazine；

3-{ [3-(1-benzofuran-2-base) imidazo [1,2-b] pyridazine-6-base] epoxide }-N-(propane-2-base) propane-1- Amine；

N-(2-{ [3-(1-benzofuran-2-base) imidazo [1,2-b] pyridazine-6-base] epoxide } ethyl) propane-2-amine；

3-(1-benzofuran-2-base)-6-[(2S)-morpholine-2-ylmethoxy] imidazo [1,2-b] pyridazine；

N-(2-{ [3-(1-benzofuran-2-base) imidazo [1,2-b] pyridazine-6-base] epoxide } ethyl)-2,2-dimethyl propylene Alkane-1-amine；

3-(1-benzofuran-2-base)-6-{2-[(3S)-morpholine-3-base] ethyoxyl } imidazo [1,2-b] pyridazine；

6-(azetidine-3-ylmethoxy)-3-(1-benzofuran-2-base) imidazo [1,2-b] pyridazine；

3-(1-benzofuran-2-base)-6-{2-[(2S)-pyrrolidin-2-yl] ethyoxyl } imidazo [1,2-b] pyridazine；

3-(1-benzofuran-2-base)-6-(piperidin-2-yl methoxyl group) imidazo [1,2-b] pyridazine；

N-(2-{ [3-(1-benzofuran-2-base) imidazo [1,2-b] pyridazine-6-base] epoxide } ethyl) cyclopropylamine；

N-(2-{ [3-(1-benzofuran-2-base) imidazo [1,2-b] pyridazine-6-base] epoxide } ethyl)-2-methylpropane-2- Amine；

2-{ [3-(1-benzofuran-2-base) imidazo [1,2-b] pyridazine-6-base] epoxide }-N-(propane-2-base) propane-1- Amine；

3-(5-chloro-1-benzofuran-2-base)-6-[(2R)-morpholine-2-ylmethoxy] imidazo [1,2-b] pyridazine；

3-(1-benzofuran-2-base)-6-[(2R)-pyrrolidin-2-yl methoxyl group] imidazo [1,2-b] pyridazine；

3-(1-benzofuran-2-base)-6-(piperidines-3-base epoxide) imidazo [1,2-b] pyridazine；

3-(1-benzofuran-2-base)-6-[(2S)-pyrrolidin-2-yl methoxyl group] imidazo [1,2-b] pyridazine；

1-{ [3-(1-benzofuran-2-base) imidazo [1,2-b] pyridazine-6-base] epoxide }-N-methylpropane-2-amine；

3-(5-chloro-1-benzofuran-2-base)-6-{2-[(2S)-pyrrolidin-2-yl] ethyoxyl } imidazo [1,2-b] pyridazine；

2-{ [3-(1-benzofuran-2-base) imidazo [1,2-b] pyridazine-6-base] epoxide }-N-methylpropane-1-amine；

Formic acid-N-(2-{ [3-(1-benzofuran-2-base) imidazo [1,2-b] pyridazine-6-base] epoxide }-2-phenylethyl) third Alkane-2-amine；

N-(2-{ [3-(1-benzofuran-2-base) imidazo [1,2-b] pyridazine-6-base] epoxide }-2-phenylethyl)-2-methyl Propane-1-amine；

(-)-2-{ [3-(1-benzofuran-2-base) imidazo [1,2-b] pyridazine-6-base] epoxide }-N-methyl-2-phenyl second Amine；

N-(2-{ [3-(1-benzofuran-2-base) imidazo [1,2-b] pyridazine-6-base] epoxide }-2-phenylethyl)-2,2-two Methylpropane-1-amine；

3-(1-benzofuran-2-base)-6-[(3R)-pyrrolidin-3-yl epoxide] imidazo [1,2-b] pyridazine；

3-(1-benzofuran-2-base)-6-[(3R)-piperidines-3-base epoxide] imidazo [1,2-b] pyridazine；

3-(4-fluoro-1-benzofuran-2-base)-6-[(2R)-morpholine-2-ylmethoxy] imidazo [1,2-b] pyridazine；

With

3-(5-fluoro-1-benzofuran-2-base)-6-[(2R)-morpholine-2-ylmethoxy] imidazo [1,2-b] pyridazine.

7. the method for the compound of the preparation logical formula (I) any one of claim 1-6, described method includes logical formula V Midbody compound:

Wherein A, R2, R3, R4 and n compound of mutual-through type (I) as any one of claim 1-6 defines, and X represent from Remove group,

The step reacted with the compound of logical formula (III):

Wherein R1 and R5 compound of mutual-through type (I) as any one of claim 1-6 defines,

Thus obtain the compound of logical formula (I):

Wherein A, R1, R2, R3, R4, R5 and n compound of mutual-through type (I) as any one of claim 1-6 defines.

8. the method for claim 7, wherein said leaving group is halogen atom or perfluoroalkyl sulfonate ester base.

9. the method for claim 8, wherein said halogen atom is chlorine atom, bromine atoms or atomic iodine.

10. the method for claim 8, wherein said perfluoroalkyl sulfonate ester base is trifluoromethane sulfonic acid ester group or nine fluorine butyl sulphurs Perester radical.

11. pharmaceutical compositions, its compound comprising logical formula (I) any one of claim 1-6 or its stereoisomer, mutually Tautomeric or pharmaceutically acceptable salt, or their mixture, and pharmaceutically acceptable diluent or carrier.

12. drug regimens, it comprises:

-one or more are selected from the first active component of the compound of logical formula (I) any one of claim 1-6, and

-one or more are selected from the second active component of chemotherapeutic anti-cancer agent.

The compound of the logical formula (I) any one of 13. claim 1-6 or its stereoisomer, tautomer or pharmaceutically Acceptable salt, or their mixture is for preparing the purposes of the medicine of prevention or treatment disease.

The purposes of 14. claim 13, wherein said disease be uncontrolled cell growth, breed and/or survive, inappropriate Cellullar immunologic response or the disease that causes of unsuitable cellular inflammation response.

The purposes of 15. claim 14, wherein said uncontrolled cell growth, breed and/or survive, unsuitable cell Immunne response or unsuitable cellular inflammation response are mediated by MKNK-1 path.

16. the purposes of claim 14, wherein said uncontrolled cell growth, breed and/or survive, unsuitable cell The disease that immunne response or unsuitable cellular inflammation response cause is neoplastic hematologic disorder, solid tumor and/or their transfer.

The purposes of 17. claim 14, wherein said uncontrolled cell growth, breed and/or survive, unsuitable cell The disease that immunne response or unsuitable cellular inflammation response cause is selected from leukemia and myelodysplastic syndrome, pernicious pouring Bar tumor, head and tumor colli including cerebroma and brain metastes, include that non-fire power and small cell lung tumor exist In breast tumor, gastroenteric tumor, endocrine tumors, breast tumor and other gynecological tumor, include tumor of kidney, tumor of bladder With tumor of prostate in interior urologic neoplasms, cutaneous tumor and sarcoma, and/or their transfer.

18. lead to the compound of formula V for preparing the purposes of the compound of the logical formula (I) any one of claim 1-6:

Wherein A, R2, R3, R4 and n compound of mutual-through type (I) as any one of claim 1-6 defines, and X represent from Remove group.

The purposes of 19. claim 18, wherein said leaving group is halogen atom or perfluoroalkyl sulfonate ester base.

The purposes of 20. claim 19, wherein said halogen atom is chlorine atom, bromine atoms or atomic iodine.

The purposes of 21. claim 19, wherein said perfluoroalkyl sulfonate ester base is trifluoromethane sulfonic acid ester group or nine fluorine butyl Sulfonate group.